Formulations for treating ocular diseases and conditions

ABSTRACT

Diseases and conditions associated with tissues of the body, including but not limited to tissues in the eye, can be effectively treated, prevented, inhibited, onset delayed, or regression caused by administering therapeutic agents to those tissues. Described herein are liquid formulations which deliver a variety of therapeutic agents, including but not limited to rapamycin, to a subject for an extended period of time; liquid formulations which form a non-dispersed mass when placed in an aqueous medium of a subject; non-dispersed mass-forming liquid formulations which form a gel or gel-like substance in an aqueous medium; liquid formulations, comprising a therapeutic agent and a plurality of polymers; and methods for delivering therapeutic agents to a subject for an extended period of time using the liquid formulations. The liquid formulation may be placed in an aqueous medium of a subject, including but not limited to via intraocular or periocular administration, or placement proximate to a site of a disease or condition to be treated in a subject. A method may be used to administer rapamycin to treat or prevent angiogenesis, choroidal neovascularization, or age-related macular degeneration, or wet age-related macular degeneration in a subject. The liquid formulations may comprise rapamycin or other therapeutic agents.

This application is a Continuation-In-Part of U.S. application Ser. No.11/351,761, filed Feb. 9, 2006, which claims benefit of ProvisionalApplication No. 60/651,790, filed Feb. 9, 2005, Provisional ApplicationNo. 60/664,040, filed Mar. 21, 2005, and Provisional Application No.60/664,306, filed Mar. 21, 2005. This application is also aContinuation-In-Part of U.S. application Ser. No. 11/352,092, filed Feb.9, 2006, which claims benefit of Provisional Application No. 60/651,790,filed Feb. 9, 2005, Provisional Application No. 60/664,040, filed Mar.21, 2005, and Provisional Application No. 60/664,306, filed Mar. 21,2005. In addition this application is a Continuation-In-Part of U.S.application Ser. No. 11/386,290, filed Mar. 21, 2006, which claimsbenefit of Provisional Application No. 60/664,119, filed Mar. 21, 2005,and Provisional Application No. 60/666,872, filed Mar. 30, 2005. ThisApplication claims benefit of Provisional Application No. 60/965,282,filed Aug. 16, 2007, and Provisional Application No. 60/965,258, filedAug. 16, 2007.

FIELD

Described herein are liquid formulations for treatment, prevention,inhibition, delaying onset of, or causing regression of a disease orcondition by delivery of therapeutic agents to a subject, including butnot limited to a human subject, including but not limited to thetreatment of age-related macular degeneration (AMD) and macular edema bydelivery of a liquid formulation comprising a therapeutic agent,including but not limited to rapamycin (sirolimus), to the eye of asubject, including but not limited to a human subject. Nonlimitingexamples of liquid formulations include solutions, suspensions, and insitu gelling formulations.

BACKGROUND

The retina of the eye contains the cones and rods that detect light. Inthe center of the retina is the macula lutea, which is about ⅓ to ½ cmin diameter. The macula provides detailed vision, particularly in thecenter (the fovea), because the cones are higher in density. Bloodvessels, ganglion cells, inner nuclear layer and cells, and theplexiform layers are all displaced to one side (rather than restingabove the cones), thereby allowing light a more direct path to thecones.

Under the retina are the choroid, comprising a collection of bloodvessels embedded within a fibrous tissue, and the deeply pigmentedepithelium, which overlays the choroid layer. The choroidal bloodvessels provide nutrition to the retina (particularly its visual cells).

There are a variety of retinal disorders for which there is currently notreatment or for which the current treatment is not optimal. Retinaldisorders such as uveitis (an inflammation of the uveal tract: iris,ciliary body, and choroid), central retinal vein occlusive diseases(CRVO), branch retinal venous occlusion (BRVO), macular degeneration,macular edema, proliferative diabetic retinopathy, and retinaldetachment generally are all retinal disorders that are difficult totreat with conventional therapies.

Age-related macular degeneration (AMD) is the major cause of severevisual loss in the United States for individuals over the age of 60. AMDoccurs in either an atrophic or less commonly an exudative form. Theatrophic form of AMD is also called “dry AMD,” and the exudative form ofAMD is also called “wet AMD.”

In exudative AMD, blood vessels grow from the choriocapillaris throughdefects in Bruch's membrane, and in some cases the underlying retinalpigment epithelium. Organization of serous or hemorrhagic exudatesescaping from these vessels results in fibrovascular scarring of themacular region with attendant degeneration of the neuroretina,detachment and tears of the retinal pigment epithelium, vitreoushemorrhage and permanent loss of central vision. This process isresponsible for more than 80% of cases of significant visual loss insubjects with AMD. Current or forthcoming treatments include laserphotocoagulation, photodynamic therapy, treatment with VEGF antibodyfragments, treatment with pegylated aptamers, and treatment with certainsmall molecule agents.

Several studies have recently described the use of laserphotocoagulation in the treatment of initial or recurrent neovascularlesions associated with AMD (Macular Photocoagulation Study Groups(1991) in Arch. Ophthal. 109:1220; Arch. Ophthal. 109:1232; Arch.Ophthal. 109:1242). Unfortunately, AMD subjects with subfoveal lesionssubjected to laser treatment experienced a rather precipitous reductionin visual acuity (mean 3 lines) at 3 months follow-up. Moreover, at twoyears post-treatment treated eyes had only marginally better visualacuity than their untreated counterparts (means of 20/320 and 20/400,respectively). Another drawback of the procedure is that vision aftersurgery is immediately worse.

Photodynamic therapy (PDT) is a form of phototherapy, a termencompassing all treatments that use light to produce a beneficialreaction in a subject. Optimally, PDT destroys unwanted tissue whilesparing normal tissue. Typically, a compound called a photosensitizer isadministered to the subject. Usually, the photosensitizer alone haslittle or no effect on the subject. When light, often from a laser, isdirected onto a tissue containing the photosensitizer, thephotosensitizer is activated and begins destroying targeted tissue.Because the light provided to the subject is confined to a particularlytargeted area, PDT can be used to selectively target abnormal tissue,thus sparing surrounding healthy tissue. PDT is currently used to treatretinal diseases such as AMD. PDT is currently the mainstay of treatmentfor subfoveal choroidal neovascularization in subjects with AMD(Photodynamic Therapy for Subfoveal Choroidal Neovascularization in AgeRelated Macular Degeneration with Verteporfin (TAP Study Group) ArchOpthalmol. 1999 117:1329-1345.

Choroidal neovascularization (CNV) has proven to be recalcitrant totreatment in most cases. Conventional laser treatment can ablate CNV andhelp to preserve vision in selected cases not involving the center ofthe retina, but this is limited to only about 10% of the cases.Unfortunately, even with successful conventional laser photocoagulation,the neovascularization recurs in about 50-70% of eyes (50% over 3 yearsand >60% at 5 years). (Macular Photocoagulation Study Group, Arch.Opthalmol. 204:694-701 (1986)). In addition, many subjects who developCNV are not good candidates for laser therapy because the CNV is toolarge for laser treatment, or the location cannot be determined so thatthe physician cannot accurately aim the laser. Photodynamic therapy,although utilized in up to 50% of new cases of subfoveal CNV has onlymarginal benefits over natural history, and generally delays progressionof visual loss rather than improving vision which is already decreasedsecondary to the subfoveal lesion. PDT is neither preventive ordefinitive. Several PDT treatments are usually required per subject andadditionally, certain subtypes of CNV fare less well than others.

Thus, there remains a long-felt need for methods, compositions, andformulations that may be used to optimally prevent or significantlyinhibit choroidal neovascularization and to prevent and treat wet AMD.

In addition to AMD, choroidal neovascularization is associated with suchretinal disorders as presumed ocular histoplasmosis syndrome, myopicdegeneration, angioid streaks, idiopathic central serouschorioretinopathy, inflammatory conditions of the retina and or choroid,and ocular trauma. Angiogenic damage associated with neovascularizationoccurs in a wide range of disorders including diabetic retinopathy,venous occlusions, sickle cell retinopathy, retinopathy of prematurity,retinal detachment, ocular ischemia and trauma.

Uveitis is another retinal disorder that has proven difficult to treatusing existing therapies. Uveitis is a general term that indicates aninflammation of any component of the uveal tract. The uveal tract of theeye consists of the iris, ciliary body, and choroid. Inflammation of theoverlying retina, called retinitis, or of the optic nerve, called opticneuritis, may occur with or without accompanying uveitis.

Uveitis is most commonly classified anatomically as anterior,intermediate, posterior, or diffuse. Posterior uveitis signifies any ofa number of forms of retinitis, choroiditis, or optic neuritis. Diffuseuveitis implies inflammation involving all parts of the eye, includinganterior, intermediate, and posterior structures.

The symptoms and signs of uveitis may be subtle, and vary considerablydepending on the site and severity of the inflammation. Regardingposterior uveitis, the most common symptoms include the presence offloaters and decreased vision. Cells in the vitreous humor, white oryellow-white lesions in the retina and/or underlying choroid, exudativeretinal detachments, retinal vasculitis, and optic nerve edema may alsobe present in a subject suffering from posterior uveitis.

Ocular complications of uveitis may produce profound and irreversibleloss of vision, especially when unrecognized or treated improperly. Themost frequent complications of posterior uveitis include retinaldetachment; neovascularization of the retina, optic nerve, or iris; andcystoid macular edema.

Macular edema (ME) can occur if the swelling, leaking, and hard exudatesnoted in background diabetic retinopathy (BDR) occur within the macula,the central 5% of the retina most critical to vision. Backgrounddiabetic retinopathy (BDR) typically consists of retinal microaneurismsthat result from changes in the retinal microcirculation. Thesemicroaneurisms are usually the earliest visible change in retinopathyseen on exam with an opthalmoscope as scattered red spots in the retinawhere tiny, weakened blood vessels have ballooned out. The ocularfindings in background diabetic retinopathy progress to cotton woolspots, intraretinal hemorrhages, leakage of fluid from the retinalcapillaries, and retinal exudates. The increased vascular permeabilityis also related to elevated levels of local growth factors such asvascular endothelial growth factor. The macula is rich in cones, thenerve endings that detect color and upon which daytime vision depends.When increased retinal capillary permeability effects the macula,blurring occurs in the middle or just to the side of the central visualfield, rather like looking through cellophane. Visual loss may progressover a period of months, and can be very annoying because of theinability to focus clearly. ME is a common cause of severe visualimpairment.

There have been many attempts to treat CNV and its related diseases andconditions, as well as other conditions such as macular edema andchronic inflammation, with pharmaceuticals. For example, use ofrapamycin to inhibit CNV and wet AMD has been described in U.S.application Ser. No. 10/665,203, which is incorporated herein byreference in its entirety. The use of rapamycin to treat inflammatorydiseases of the eye has been described in U.S. Pat. No. 5,387,589,titled Method of Treating Ocular Inflammation, with inventor PrassadKulkarni, assigned to University of Louisville Research Foundation, thecontents of which is incorporated herein in its entirety.

Particularly for chronic diseases, including those described herein,there is a great need for long acting methods for delivering therapeuticagents to the eye, such as to the posterior segment to treat CNV in suchdiseases as AMD, macular edema, proliferative retinopathies, and chronicinflammation. Formulations with extended delivery of therapeutic agentare more comfortable and convenient for a subject, due to a diminishedfrequency of ocular injections of the therapeutic agent.

Direct delivery of therapeutic agents to the eye rather than systemicadministration may be advantageous because the therapeutic agentconcentration at the site of action is increased relative to thetherapeutic agent concentration in a subject's circulatory system.Additionally, therapeutic agents may have undesirable side effects whendelivered systemically to treat posterior segment disease. Thus,localized drug delivery may promote efficacy while decreasing sideeffects and systemic toxicity.

SUMMARY

The methods, compositions, and liquid formulations described hereinallow delivery of a therapeutic agent to a subject, including but notlimited to a human subject or to the eye(s) of a subject. Describedherein are methods, compositions, and liquid formulations for deliveringa variety of therapeutic agents for extended periods of time which canbe used for the treatment, prevention, inhibition, delaying onset of, orcausing regression of a number of conditions or diseases, including butnot limited to diseases or conditions of the eye. The liquidformulations include, without limitation, solutions, suspensions, and insitu gelling formulations.

Described herein are methods, compositions and liquid formulations foradministering to a human subject an amount of rapamycin effective totreat, prevent, inhibit, delay onset of, or cause regression of wet AMD,dry AMD, or macular edema (including, without limitation, diabeticmacular edema).

As described in further detail in the Detailed Description section, themethods, compositions and liquid formulations may also be used fordelivery to a subject, including but not limited to a human subject orto the eye(s) of a human subject of therapeutically effective amounts ofrapamycin for the treatment, prevention, inhibition, delaying of theonset of, or causing the regression of wet AMD, dry AMD, or macularedema. In some variations, the methods, compositions, and liquidformulations are used to treat wet AMD. In some variations, the methods,compositions, and liquid formulations are used to prevent wet AMD. Insome variations, the methods, compositions, and liquid formulations areused to treat dry AMD. In some variations, the methods, compositions,and liquid formulations are used to prevent dry AMD. In some variations,the methods and formulations described herein are used to prevent thetransition from dry AMD to wet AMD. In some variations, the methods andformulations described herein are used to treat macular edema(including, without limitation, diabetic macular edema). In somevariations, the methods and formulations described herein are used toprevent macular edema. The methods, compositions and liquid formulationsmay also be used for delivery to a subject, including but not limited toa human subject or to the eye of a subject of therapeutically effectiveamounts of rapamycin for the treatment, prevention, inhibition, delayingof the onset of, or causing the regression of CNV. In some variations,the methods, compositions and liquid formulations are used to treat CNV.The methods, compositions and liquid formulations may also be used fordelivery to a subject, including but not limited to a human subject orto the eye of a subject of therapeutically effective amounts ofrapamycin for the treatment, prevention, inhibition, delaying of theonset of, or causing the regression of angiogenesis in the eye. In somevariations, the methods, compositions and liquid formulations are usedto treat angiogenesis. Other diseases and conditions that may betreated, prevented, inhibited, have onset delayed, or caused to regressusing rapamycin are described in the Diseases and Conditions section ofthe Detailed Description.

As described in further detail in the Detailed Description, the methods,compositions and liquid formulations may also be used for delivery to asubject, including but not limited to a human subject or to the eye of asubject of therapeutically effective amounts of therapeutic agents otherthan rapamycin for the treatment, prevention, inhibition, delaying ofthe onset of, or causing the regression of wet AMD or macular edema. Insome variations, the methods, compositions and liquid formulations areused to treat wet AMD. In some variations, the methods, compositions andliquid formulations are used to treat macular edema. Therapeutic agentsthat may be used are described in detail in the Therapeutic Agentssection. Such therapeutic agents include but are not limited toimmunophilin binding compounds. Immunophilin binding compounds that maybe used include but are not limited to the limus family of compoundsdescribed further in the Therapeutic Agents section herein, includingrapamycin, SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779,AP23841, ABT-578, derivatives, analogs, prodrugs, salts and estersthereof. In certain variations, the therapeutic agent is dasatinib, or asalt or ester thereof. The methods, compositions and liquid formulationsmay also be used for delivery to a subject, including but not limited toa human subject or to the eye of a subject of therapeutically effectiveamounts of therapeutic agents for the treatment, prevention, inhibition,delaying of the onset of, or causing the regression of CNV. In somevariations, the methods, compositions and liquid formulations are usedto treat CNV. The methods, compositions and liquid formulations may alsobe used for delivery to a subject, including but not limited to a humansubject or to the eye of a subject of therapeutically effective amountsof therapeutic agents for the treatment, prevention, inhibition,delaying of the onset of, or causing the regression of angiogenesis inthe eye. In some variations, the methods, compositions and liquidformulations are used to treat angiogenesis. Other diseases andconditions that may be treated, prevented, inhibited, have onsetdelayed, or caused to regress using therapeutic agents other thanrapamycin are described in the Diseases and Conditions section of theDetailed Description.

One liquid formulation described herein comprises a solution thatincludes a therapeutic agent dissolved in a solvent. Generally, anysolvent that has the desired effect may be used in which the therapeuticagent dissolves and which can be administered to a subject, includingbut not limited to a human subject or an eye of a subject. Generally,any concentration of therapeutic agent that has the desired effect canbe used. The formulation in some variations is a solution which isunsaturated, a saturated or a supersaturated solution. The solvent maybe a pure solvent or may be a mixture of liquid solvent components. Insome variations the solution formed is an in situ gelling formulation.Solvents and types of solutions that may be used are well known to thoseversed in such drug delivery technologies.

The liquid formulations described herein may form a non-dispersed masswhen placed into a rabbit eye, including but not limited to the vitreousof a rabbit eye. In some variations the non-dispersed mass comprises agel. In some variations, the liquid formulation comprises a therapeuticagent and a plurality of polymers. In some variations one of thepolymers is polyacrylate or polymethacrylate. In some variations one ofthe polymers is polyvinylpyrrolidone.

The liquid formulations described herein may form a non-dispersed masswhen placed into a human eye, including, but not limited to, injectedinto the vitreous of a human eye or injected between the sclera andconjunctiva of a human eye.

In some variations, the non-dispersed mass comprises a depot. In somevariations, the non-dispersed mass consists of a depot.

For liquid formulations which form a non-dispersed mass, thenon-dispersed mass may generally be any geometry or shape. Thenon-dispersed mass-forming liquid formulations may, for instance, appearas a compact spherical mass when placed in the vitreous. In somevariations the liquid formulations described herein form a milky orwhitish colored semi-contiguous or semi-solid non-dispersed massrelative to the medium in which it is placed, when placed in thevitreous.

The liquid formulations may generally be administered in any volume thathas the desired effect. In one method a volume of a liquid formulationis administered to the vitreous and the liquid formulation is less thanone half the volume of the vitreous.

Routes of administration that may be used to administer a liquidformulation include but are not limited to (1) placement of the liquidformulation by placement, including by injection, into a medium,including but not limited to an aqueous medium in the body, includingbut not limited to intraocular or periocular injection; or (2) oraladministration of the liquid formulation. The liquid formulation may beadministered systemically, including but not limited to the followingdelivery routes: rectal, vaginal, infusion, intramuscular,intraperitoneal, intraarterial, intrathecal, intrabronchial,intracisternal, cutaneous, subcutaneous, intradermal, transdermal,intravenous, intracervical, intraabdominal, intracranial,intrapulmonary, intrathoracic, intratracheal, nasal, buccal, sublingual,oral, parenteral, or nebulised or aerosolized using aerosol propellants.In some variations, the liquid formulation is administeredsubconjunctivally. In some variations, the liquid formulation isadministered intravitreally. In some alternative variations, the liquidformulation is administered by subtenon injection. In some variations,the liquid formulation is administered topically.

The liquid formulations described herein may be delivered to any mediumof a subject, including but not limited to a human subject, includingbut not limited to an aqueous medium of a subject.

One liquid formulation described herein comprises a liquid formulationof rapamycin or other therapeutic agent. The liquid formulations maycomprise a solution, suspension, an in situ gelling formulation, or anemulsion. The droplets in the emulsion may generally be of any size,including but not limited to up to about 5,000 nm.

In some formulations described herein, the liquid formulations maycomprise a therapeutic agent including but not limited to rapamycin, andone or more solubilizing agents or solvents. In some variations, thesolubilizing agent or solvent is glycerin, DMSO, DMA,N-methylpyrrolidone, ethanol, benzyl alcohol, isopropyl alcohol,polyethylene glycol of various molecular weights, including but notlimited to PEG 300 and PEG 400, or propylene glycol or a mixture of oneor more thereof.

In some formulations described herein, the liquid formulation includeshyaluronic acid.

The liquid formulations described herein may deliver a therapeutic agentor agents for an extended period of time. One nonlimiting example ofsuch an extended release delivery system is a liquid formulation thatdelivers a therapeutic agent or agents to a subject, including but notlimited to a human subject or to the eye of a subject in an amountsufficient to maintain an amount effective to treat, prevent, inhibit,delay onset of, or cause regression of a disease or condition in asubject for an extended period of time. In some variations, the liquidformulation is used to treat a disease or condition in a subject,including but not limited to a human subject. In some variations, theliquid formulation delivers the therapeutic agent for at least aboutone, about two, about three, about six, about nine, or about twelvemonths.

The liquid formulations described herein may deliver rapamycin or othertherapeutic agents for an extended period of time. One nonlimitingexample of such an extended release delivery system is a liquidformulation that delivers rapamycin to a subject, including but notlimited to a human subject or to the eye of a subject in an amountsufficient to maintain an amount effective to treat, prevent, inhibit,delay onset of, or cause regression of wet age-related maculardegeneration for an extended period of time. In some variations, theliquid formulation is used to treat wet age-related macular degenerationfor an extended period of time. In some variations, the liquidformulation is used to prevent wet age-related macular degeneration foran extended period of time. In some variations, the liquid formulationis used to prevent transition of dry AMD to wet AMD for an extendedperiod of time. In one nonlimiting example, the liquid formulationdelivers the rapamycin to the vitreous, sclera, retina, choroid, macula,or other tissues of a subject, including but not limited to a humansubject in an amount sufficient to treat, prevent, inhibit, delay onsetof, or cause regression of wet age-related macular degeneration for atleast about three, about six, about nine, or about twelve months. Insome variations, the level of rapamycin is sufficient to treat AMD. Insome variations, the level of rapamycin is sufficient to prevent onsetof wet AMD.

Other extended periods of release are described in the DetailedDescription.

Described herein is a liquid formulation comprising a therapeutic agent,wherein the liquid formulation contains less than about 90% (w/w) of anymaterial having a hygroscopicity that is about equal to or greater thanthat of polyethylene glycol (PEG 400). In some variations, the liquidformulation contains less than about 80%, less than about 70%, less thanabout 60%, less than about 50%, or less than about 40% of the materialhaving a hygroscopicity that is about equal to or greater than that ofpolyethylene glycol (PEG 400). In some variations, the material is asolvent. Methods are provided for treating an ocular disease in a humansubject comprising administering to the subject by intraocular orperiocular delivery a volume of the liquid formulation containing anamount of the therapeutic agent effective to treat an ocular disease inthe subject. Methods for preventing an ocular disease in a human subjectcomprising administering to the subject by intraocular or perioculardelivery a volume of the liquid formulation containing an effectiveamount of the therapeutic agent are also provided.

Also described herein is a method for treating an ocular disease in ahuman subject comprising administering to the human subject a volume ofa liquid formulation by placement between the sclera and conjunctiva ofthe subject, wherein the liquid formulation comprises: (a) an effectiveamount of a therapeutic agent; and (b) a material selected from thegroup consisting of polyethylene glycol 400 (PEG 400), a material aboutas hygroscopic as PEG 400, or a material more hygroscopic than PEG 400.In certain variations, the liquid formulation comprises less than about80 μl, less than about 60 μl, or less than about 40 μl of such amaterial. In some variations, the material is a solvent.

Also described herein is a method for preventing an ocular disease in ahuman subject comprising administering to the human subject a volume ofa liquid formulation by placement between the sclera and conjunctiva ofthe subject, wherein the liquid formulation comprises: (a) an effectiveamount of a therapeutic agent; and (b) a material selected from thegroup consisting of polyethylene glycol 400 (PEG 400), a material aboutas hygroscopic as PEG 400, or a material more hygroscopic than PEG 400.In certain variations, the liquid formulation comprises less than about80 μl, less than about 60 μl, or less than about 40 μl of such amaterial. In some variations, the material is a solvent.

Further described herein is a method for treating an ocular disease in ahuman subject, the method comprising administering periocularly to thesubject a volume of formulation comprising an amount of therapeuticagent and a volume of an excipient, wherein the amount of therapeuticagent is sufficient to treat the ocular disease, and wherein theexcipient is PEG 400 or a material having a hygroscopicity about equalto or less than PEG 400 and the volume of the excipient is less than orequal to about 100 μl. In some variations, the volume of the excipientis less than or equal to about 80 μl, less than or equal to about 60 μl,or less than or equal to about 40 μl. In some variations, the volume offormulation is administered between the sclera and conjunctiva of thesubject. In some variations, the ocular disease is dry age-relatedmacular degeneration, wet age-related macular degeneration or macularedema. In some variations, the excipient is a solvent. In somevariations, the excipient is PEG 400.

Further described herein is a method for preventing an ocular disease ina human subject, the method comprising administering periocularly to thesubject a volume of formulation comprising an amount of therapeuticagent and a volume of an excipient, P wherein the amount of therapeuticagent is sufficient to prevent the ocular disease, and wherein theexcipient is PEG 400 or a material having a hygroscopicity about equalto or less than PEG 400 and the volume of the excipient is less than orequal to about 100 μl. In some variations, the volume of the excipientis less than or equal to about 80 μl, less than or equal to about 60 μl,or less than or equal to about 40 μl. In some variations, the volume offormulation is administered between the sclera and conjunctiva of thesubject. In some variations, the ocular disease is dry age-relatedmacular degeneration, wet age-related macular degeneration or macularedema. In some variations, the excipient is a solvent. In somevariations, the excipient is PEG 400.

Described herein is also a liquid formulation comprising a therapeuticagent, water, and one or more excipients selected from the groupconsisting of N-methylpyrrolidone (NMP), dimethyl acetamine (DMA),dimethyl sulfoxide (DMSO), propylene glycol (PG), polyethylene glycol600 (PEG 600), and polyethylene glycol 400. In some variations, theexcipient is a solvent. In certain variations, the formulation comprisesat least about five percent (w/w) of water. In some variations, inaddition to the therapeutic agent and water, the formulation comprisesboth (i) an excipient selected from the group consisting of N-methylpyrrolidone (NMP), dimethyl acetamine (DMA), and dimethyl sulfoxide(DMSO); and (ii) an excipient selected from the group consisting ofpropylene glycol (PG), polyethylene glycol 600 (PEG 600), andpolyethylene glycol 400. The formulation, may, in some variations,comprise up to about five percent (w/w) therapeutic agent. In somevariations, the formulation further comprises ethanol. Methods areprovided for treating an ocular disease in a human subject comprisingadministering to the subject by intraocular or periocular delivery avolume of the liquid formulation containing an amount of the therapeuticagent effective to treat the ocular disease in the subject. Methods forpreventing an ocular disease in a human subject comprising administeringto the subject by intraocular or periocular delivery a volume of theliquid formulation containing an effective amount of the therapeuticagent are also provided.

In some variations of each of the aforementioned methods and/orformulations, as well as other methods and/or formulations describedherein, the liquid formulation, when injected between the sclera andconjunctiva of a rabbit eye delivers an amount of the therapeutic agentsufficient to achieve, for a period of time of at least 30 daysfollowing administration of the liquid formulation, one or both of thefollowing: (a) an average concentration of therapeutic agent in thevitreous of the rabbit eye equivalent to a rapamycin concentration of atleast 0.01 ng/ml; or (b) an average concentration of therapeutic agentin the retina choroid tissues of the rabbit eye equivalent to arapamycin concentration of at least 0.001 ng/mg.

In some variations of each of the aforementioned methods and/orformulations, as well as other methods and/or formulations describedherein, the liquid formulation, when injected into the vitreous of arabbit eye delivers an amount of the therapeutic agent sufficient toachieve, for a period of time of at least 30 days followingadministration of the liquid formulation, one or both of the following:(a) an average concentration of therapeutic agent in the retina choroidtissues of the rabbit eye equivalent to a rapamycin concentration of atleast 0.01 ng/mg; or (b) an average concentration of therapeutic agentin the vitreous of the rabbit eye equivalent to a rapamycinconcentration of at least 1000 ng/ml.

Described herein is a liquid formulation comprising a therapeutic agent,wherein the liquid formulation when administered by subtenon injectionto a rabbit eye delivers an amount of the therapeutic agent sufficientto achieve, for a period of time of at least 30 days followingadministration of the liquid formulation, one or both of the following:(a) an average concentration of therapeutic agent in the vitreous of therabbit eye equivalent to a rapamycin concentration of at least 0.01ng/ml; or (b) an average concentration of therapeutic agent in theretina choroid tissues of the rabbit eye equivalent to a rapamycinconcentration of at least 0.001 ng/mg. Methods are provided for treatingan ocular disease in a human subject comprising administering to thesubject by intraocular or periocular delivery a volume of the liquidformulation containing an amount of the therapeutic agent effective totreat an ocular disease in the subject. Methods for preventing an oculardisease in a human subject comprising administering to the subject byintraocular or periocular delivery a volume of the liquid formulationcontaining an effective amount of the therapeutic agent are alsoprovided.

In some variations of each of the aforementioned methods and/orformulations, as well as other methods and/or formulations describedherein, the liquid formulation is administered by intravitreal,subconjunctival, topical, or subtenon administration.

Also described herein is a liquid formulation comprising: (a) atherapeutic agent; (b) a first component selected from the groupconsisting of ethoxlyated p-tert-octylphenol formaldehyde polymer,polyoxyl 35 castor oil, propylene glycol monolaurate, propylene glycoldicaprylate/dicaprate, and macrogol 15 hydroxystearate; and (c) a secondcomponent comprising monoglycerides and/or diglycerides of caprylate. Insome variations, the formulation further comprises ethanol and/or water.Methods for treating an ocular disease in a human subject, comprisingadministering to the subject by topical delivery a volume of the liquidformulation containing an amount of the therapeutic agent effective totreat the ocular disease in the human subject are also provided. Methodsfor preventing an ocular disease in a human subject, comprisingadministering to the subject by topical delivery a volume of the liquidformulation containing an amount of the therapeutic agent effective toprevent the ocular disease in the human subject are also provided. Insome variations, the liquid formulation is applied topically to an eyeof the subject about once or less a day, about once or less every 5days, or about once or less every 10 days. In some variations, themethods are methods of treatment. In some variations, the disease to betreated is a disease of the retina. In some variations, the disease isAMD or macular edema.

In addition, described herein is a method for treating an ocular diseasein a human subject, the method comprising administering to the humansubject by topical delivery a volume of a liquid formulation containingan amount of a therapeutic agent effective to treat the ocular diseasein the human subject, wherein the liquid formulation when topicallyapplied to a rabbit eye in a volume of 80 μl a day for six days deliversan amount of the therapeutic agent sufficient to achieve: (a) an averageconcentration of the therapeutic agent in the cornea of the rabbit eyeat day 6 equivalent to a rapamycin concentration of at least 0.01 ng/mg;or (b) an average concentration of therapeutic agent in the retinachoroid tissues of the rabbit eye at day 6 equivalent to a rapamycinconcentration of at least 0.001 ng/mg. In some variations, the liquidformulation is applied topically to an eye of the subject about once orless a day, about once or less every 5 days, or about once or less every10 days. In some variations, the method is a method of treatment. Insome variations, the liquid formulation consists essentially of about 2%(w/w) rapamycin, about 4% (w/w) ethanol, and about 94% (w/w) PEG 400.

In addition, described herein is a method for preventing an oculardisease in a human subject, the method comprising administering to thehuman subject by topical delivery a volume of a liquid formulationcontaining an amount of a therapeutic agent effective to prevent theocular disease in the human subject, wherein the liquid formulation whentopically applied to a rabbit eye in a volume of 80 μl a day for sixdays delivers an amount of the therapeutic agent sufficient to achieve:(a) an average concentration of the therapeutic agent in the cornea ofthe rabbit eye at day 6 equivalent to a rapamycin concentration of atleast 0.01 ng/mg; or (b) an average concentration of therapeutic agentin the retina choroid tissues of the rabbit eye at day 6 equivalent to arapamycin concentration of at least 0.001 ng/mg. In some variations, theliquid formulation is applied topically to an eye of the subject aboutonce or less a day, about once or less every 5 days, or about once orless every 10 days. In some variations, the method is a method oftreatment. In some variations, the liquid formulation consistsessentially of about 2% (w/w) rapamycin, about 4% (w/w) ethanol, andabout 94% (w/w) PEG 400.

Further described herein is a liquid formulation comprising atherapeutic agent and water, wherein the liquid formulation when appliedto a rabbit eye in a volume of 80 μl a day for six days delivers anamount of the therapeutic agent sufficient to achieve: (a) an averageconcentration of the therapeutic agent in the cornea of the rabbit eyeat day 6 equivalent to a rapamycin concentration of at least 0.01 ng/mg;or (b) an average concentration of therapeutic agent in the retinachoroid tissues of the rabbit eye at day 6 equivalent to a rapamycinconcentration of at least 0.001 ng/mg. Methods for treating an oculardisease in a human subject, comprising administering to the subject bytopical delivery a volume of the liquid formulation containing an amountof the therapeutic agent effective to treat the ocular disease in thehuman subject are also provided. Methods for preventing an oculardisease in a human subject, comprising administering to the subject bytopical delivery a volume of the liquid formulation containing an amountof the therapeutic agent effective to prevent the ocular disease in thehuman subject are also provided. In some variations, the liquidformulation is applied topically to an eye of the subject about once orless a day, about once or less every 5 days, or about once or less every10 days. In some variations, the methods are methods of treatment.

Also described herein is a method for treating an ocular disease in ahuman subject, the method comprising topically administering to thesubject a liquid formulation comprising a therapeutic agent, wherein theliquid formulation, when a volume of about 40 μl is applied topically toa rabbit eye, delivers an amount of the therapeutic agent sufficient toachieve: (a) an average concentration of the therapeutic agent in thecornea of the rabbit eye equivalent to an average rapamycinconcentration of at least 0.01 ng/mg for at least about ten days afteradministration of the liquid formulation; or (b) an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye equivalent to an average rapamycin concentration of at least0.001 ng/mg for at least about ten days after administration of theliquid formulation. In some variations, the liquid formulation furthercomprises polyethylene glycol. In some variations, the liquidformulation is applied topically to an eye of the subject about once orless a day, about once or less every 5 days, or about once or less every10 days. In some variations, the methods are methods of treatment. Insome variations, the liquid formulation consists essentially of about 2%(w/w) rapamycin, about 4% (w/w) ethanol, and about 94% (w/w) PEG 400.

Also described herein is a method for preventing an ocular disease in ahuman subject, the method comprising topically administering to thesubject a liquid formulation comprising a therapeutic agent, wherein theliquid formulation, when a volume of about 40 μl is applied topically toa rabbit eye, delivers an amount of the therapeutic agent sufficient toachieve: (a) an average concentration of the therapeutic agent in thecornea of the rabbit eye equivalent to an average rapamycinconcentration of at least 0.01 ng/mg for at least about ten days afteradministration of the liquid formulation; or (b) an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye equivalent to an average rapamycin concentration of at least0.001 ng/mg for at least about ten days after administration of theliquid formulation. In some variations, the liquid formulation furthercomprises polyethylene glycol. In some variations, the liquidformulation is applied topically to an eye of the subject about once orless a day, about once or less every 5 days, or about once or less every10 days. In some variations, the methods are methods of treatment. Insome variations, the liquid formulation consists essentially of about 2%(w/w) rapamycin, about 4% (w/w) ethanol, and about 94% (w/w) PEG 400.

In some variations of each of the aforementioned methods and/orformulations, as well as other methods and/or formulations describedherein, the therapeutic agent is rapamycin, SDZ-RAD, tacrolimus,everolimus, pimecrolimus, CCI-779, AP23841, or ABT-578, or is aderivative, analog, prodrug, salt or ester of rapamycin, SDZ-RAD,tacrolimus, everolimus, pimecrolimus, CCI-779, AP23841, or ABT-578.

In some variations of each of the aforementioned methods and/orformulations, as well as other methods and/or formulations describedherein, the therapeutic agent is rapamycin, or a pharmaceuticallyacceptable salt or ester thereof.

In some variations of each of the aforementioned methods and/orformulations, as well as other methods and/or formulations describedherein, the therapeutic agent is dasatinib, or a pharmaceuticallyacceptable salt or ester thereof.

Further described herein is a liquid formulation consisting essentiallyof about 2% (w/w) rapamycin, about 4% (w/w) ethanol, and about 94% (w/w)PEG 400, as well as a liquid formulation consisting essentially of about4% (w/w) rapamycin, about 4% (w/w) ethanol, and about 92% (w/w) PEG 400.A liquid formulation consisting essentially of about 1.47% (w/w)rapamycin, about 2.93% (w/w) ethanol, about 26.6% (w/w) normal saline,and about 69% (w/w) PEG 400 is also provided. Methods of using suchformulations in the treatment of an ocular disease are also provided.Methods of using such formulations in the prevention of an oculardisease are also provided. In some variations, the methods compriseadministering the liquid formulation by intravitreal, subconjunctival,topical, or subtenon administration.

A method for treating an ocular disease in a human subject, the methodcomprising administering to the human subject a volume of a liquidformulation by placement in the vitreous of the subject, wherein thevolume of the liquid formulation comprises between about 20 μg and about750 μg of rapamycin, or a pharmaceutically acceptable salt or esterthereof, is also provided herein. In some variations, the volume of theliquid formulation is about 30 μl or less, about 20 μl or less, or about10 μl or less. In certain variations, the volume of the liquidformulation contains between about 30 μg and about 200 μg of rapamycin.The liquid formulation, in some variations, consists essentially ofabout 2% (w/w) rapamycin, about 4% (w/w) ethanol, and about 94% (w/w)PEG 400.

A method for preventing an ocular disease in a human subject, the methodcomprising administering to the human subject a volume of a liquidformulation by placement in the vitreous of the subject, wherein thevolume of the liquid formulation comprises between about 20 μg and about750 μg of rapamycin, or a pharmaceutically acceptable salt or esterthereof, is also provided herein. In some variations, the volume of theliquid formulation is about 30 μl or less, about 20 μl or less, or about10 μl or less. In certain variations, the volume of the liquidformulation contains between about 30 μg and about 200 μg of rapamycin.The liquid formulation, in some variations, consists essentially ofabout 2% (w/w) rapamycin, about 4% (w/w) ethanol, and about 94% (w/w)PEG 400.

In addition, described herein is a method for treating an ocular diseasein a human subject, the method comprising administering to the humansubject a volume of a liquid formulation by placement between the scleraand conjunctiva of the subject, wherein the liquid formulation comprisesbetween about 50 μg and about 3 mg of rapamycin, or a pharmaceuticallyacceptable salt or ester thereof. In some variations, the volume of theliquid formulation is about 150 μl or less, about 100 μl or less, orabout 40 μl or less. In some variations, the volume of the liquidformulation comprises between about 300 μg and about 1000 μg ofrapamycin. In some variations, the amount of rapamycin in the volume ofthe formulation that is administered is about 400 μg to about 900 μg,about 500 μg to about 800 μg, or about 600 μg to about 700 μg ofrapamycin. In some embodiments, the volume of the formulation that isadministered to the subject is about 10 μl to about 50 μl, about 15 μlto about 45 μl, about 20 μl to about 40 μl, or about 25 μl to about 35μl. In some variations, the volume of the formulation that isadministered comprises about 440 μg of rapamycin in about 20 μl. In somevariations, the volume of the formulation that is administered comprisesabout 660 μg of rapamycin in about 30 μl. In some variations, the volumeof the formulation that is administered comprises about 880 μg ofrapamycin in about 40 μl. The liquid formulation, in some variations,consists essentially of about 2% (w/w) rapamycin, about 4% (w/w)ethanol, and about 94% (w/w) PEG 400. In some variations, the liquidformulation comprises less than about 40 μl of polyethylene glycol. Insome alternative variations, the formulation comprises less than about80 μl of polyethylene glycol (including, without limitation, PEG 400),less than about 60 μl of polyethylene glycol, or less than about 40 μlof polyethylene glycol. In some variations, the formulation comprises nopolyethylene glycol.

In addition, described herein is a method for preventing an oculardisease in a human subject, the method comprising administering to thehuman subject a volume of a liquid formulation by placement between thesclera and conjunctiva of the subject, wherein the liquid formulationcomprises between about 50 μg and about 3 mg of rapamycin, or apharmaceutically acceptable salt or ester thereof. In some variations,the volume of the liquid formulation is about 150 μl or less, about 100μl or less, or about 40 μl or less. In some variations, the volume ofthe liquid formulation comprises between about 300 μg and about 1000 μgof rapamycin. In some variations, the amount of rapamycin in the volumeof the formulation that is administered is about 400 μg to about 900 μg,about 500 μg to about 800 μg, or about 600 μg to about 700 μg ofrapamycin. In some embodiments, the volume of the formulation that isadministered to the subject is about 10 μl to about 50 μl, about 15 μlto about 45 μl, about 20 μl to about 40 μl, or about 25 μl to about 35μl. In some variations, the volume of the formulation that isadministered comprises about 440 μg of rapamycin in about 20 μl. In somevariations, the volume of the formulation that is administered comprisesabout 660 μg of rapamycin in about 30 μl. In some variations, the volumeof the formulation that is administered comprises about 880 μg ofrapamycin in about 40 μl. The liquid formulation, in some variations,consists essentially of about 2% (w/w) rapamycin, about 4% (w/w)ethanol, and about 94% (w/w) PEG 400. In some variations, the liquidformulation comprises less than about 40 μl of polyethylene glycol. Insome alternative variations, the formulation comprises less than about80 μl of polyethylene glycol (including, without limitation, PEG 400),less than about 60 μl of polyethylene glycol, or less than about 40 μlof polyethylene glycol. In some variations, the formulation comprises nopolyethylene glycol.

Further described herein is a method for treating an ocular disease in ahuman subject, the method comprising administering by subtenon injectionto the subject a volume of a liquid formulation comprising between about50 μg and about 3 mg of a therapeutic agent. In some variations, thetherapeutic agent is selected from the group consisting of rapamycin ordasatinib, or a pharmaceutically acceptable salt or ester of eitheragent. In some variations, the liquid formulation comprises greater thanabout 30 μl, greater than about 60 μl, or greater than about 80 μl ofpolyethylene glycol 400 (PEG 400), or a material having a hygroscopicityequal to or greater than PEG 400. In some variations, the material is asolvent. In some variations, the liquid formulation consists essentiallyof about 2% (w/w) rapamycin, about 4% (w/w) ethanol, and about 94% (w/w)PEG 400.

Further described herein is a method for preventing an ocular disease ina human subject, the method comprising administering by subtenoninjection to the subject a volume of a liquid formulation comprisingbetween about 50 μg and about 3 mg of a therapeutic agent. In somevariations, the therapeutic agent is selected from the group consistingof rapamycin or dasatinib, or a pharmaceutically acceptable salt orester of either agent. In some variations, the liquid formulationcomprises greater than about 30 μl, greater than about 60 μl, or greaterthan about 80 μl of polyethylene glycol 400 (PEG 400), or a materialhaving a hygroscopicity equal to or greater than PEG 400. In somevariations, the material is a solvent. In some variations, the liquidformulation consists essentially of about 2% (w/w) rapamycin, about 4%(w/w) ethanol, and about 94% (w/w) PEG 400.

Also described herein is a method for treating an ocular disease in ahuman subject comprising: (a) administering to the human subject avolume of a first liquid formulation by placement in the vitreous of thesubject, wherein the first liquid formulation comprises a firsttherapeutic agent; and (b) administering to the human subject a volumeof a second liquid formulation by placement between the sclera andconjunctiva of the subject, wherein the second liquid formulationcomprises rapamycin, or a pharmaceutically acceptable salt or esterthereof; wherein an amount of the first and second therapeutic agentseffective to treat the ocular disease is administered to the subject. Insome variations, step (b) is subsequent to step (a). In some variations,the first therapeutic agent is rapamycin, or a pharmaceuticallyacceptable salt or ester thereof. The first liquid formulation may beidentical to the second liquid formulation or different from the secondliquid formulation. In some variations, the first therapeutic agent isselected from the group consisting of ranibizumab and bevacizumab. Insome variations, both step (a) and step (b) are performed during asingle administration session (including, but not limited to, a singlevisit with a physician or other medical professional or other singlesession in which the administrations are performed by the subject, aphysician, or other medical professional). In some variations, the firstand/or second liquid formulation is a liquid formulation consistingessentially of about 2% (w/w) rapamycin, about 4% (w/w) ethanol, andabout 94% (w/w) PEG 400.

Also described herein is a method for preventing an ocular disease in ahuman subject comprising: (a) administering to the human subject avolume of a first liquid formulation by placement in the vitreous of thesubject, wherein the first liquid formulation comprises a firsttherapeutic agent; and (b) administering to the human subject a volumeof a second liquid formulation by placement between the sclera andconjunctiva of the subject, wherein the second liquid formulationcomprises rapamycin, or a pharmaceutically acceptable salt or esterthereof; wherein an amount of the first and second therapeutic agentseffective to prevent the ocular disease is administered to the subject.In some variations, step (b) is subsequent to step (a). In somevariations, the first therapeutic agent is rapamycin, or apharmaceutically acceptable salt or ester thereof. The first liquidformulation may be identical to the second liquid formulation ordifferent from the second liquid formulation. In some variations, thefirst therapeutic agent is selected from the group consisting ofranibizumab and bevacizumab. In some variations, both step (a) and step(b) are performed during a single administration session (including, butnot limited to, a single visit with a physician or other medicalprofessional or other single session in which the administrations areperformed by the subject, a physician, or other medical professional).In some variations, the first and/or second liquid formulation is aliquid formulation consisting essentially of about 2% (w/w) rapamycin,about 4% (w/w) ethanol, and about 94% (w/w) PEG 400.

In some variations of each of the aforementioned methods and/orformulations, as well as other methods and/or formulations describedherein, the liquid formulation comprises between about 0.001% (w/w) toabout 10% (w/w) therapeutic agent. In some variations, the formulationcomprises between about 0.001% (w/w) to about 1.0% (w/w) the therapeuticagent. In some variations, the formulation comprises between about 0.01%(w/w) to about 0.1% (w/w) therapeutic agent. In further variations, theformulation comprises between about 0.1% (w/w) to about 6% (w/w)therapeutic agent. In some variations, the formulation contains up toabout 5% rapamycin.

Further described herein is a method for treating an ocular disease in ahuman subject, the method comprising: (a) administering to the humansubject a volume of a first therapeutic agent; and (b) administering tothe human subject by intraocular or periocular delivery a volume of aliquid formulation (including, without limitation, by intravitreal,subconjunctival, topical, or subtenon administration), wherein theliquid formulation consists essentially of about 2% (w/w) rapamycin,about 4% (w/w) ethanol, and about 94% (w/w) PEG 400, and wherein anamount of the first therapeutic agent and rapamycin effective to treatthe ocular disease is administered to the subject. In some variations,the first therapeutic agent is selected from the group consisting ofranibizumab, bevacizumab, and verteporfin. The first therapeutic agentmay, in some variations, be administered to the subject by intraoculardelivery.

Further described herein is a method for preventing an ocular disease ina human subject, the method comprising: (a) administering to the humansubject a volume of a first therapeutic agent; and (b) administering tothe human subject by intraocular or periocular delivery a volume of aliquid formulation (including, without limitation, by intravitreal,subconjunctival, topical, or subtenon administration), wherein theliquid formulation consists essentially of about 2% (w/w) rapamycin,about 4% (w/w) ethanol, and about 94% (w/w) PEG 400, and wherein anamount of the first therapeutic agent and rapamycin effective to preventthe ocular disease is administered to the subject. In some variations,the first therapeutic agent is selected from the group consisting ofranibizumab, bevacizumab, and verteporfin. The first therapeutic agentmay, in some variations, be administered to the subject by intraoculardelivery.

Also described herein is a method for treating an ocular disease in ahuman subject, the method comprising: (a) diluting a liquid formulationconsisting essentially of about 2% (w/w) rapamycin, about 4% (w/w)ethanol, and about 94% (w/w) PEG 400 with aqueous liquid to produce adiluted liquid formulation; and (b) administering to the human subjectby intraocular or periocular delivery a volume of the diluted liquidformulation, wherein an amount of rapamycin effective to treat theocular disease is administered to the subject. In some variations, theformulation is a clear solution. In some variations, the dilution is upto about a 1.4-fold dilution. In some variations, the liquid formulationis diluted up to about 1.2 fold. In some variations, the aqueous liquidis saline. In some variations, the diluted liquid formulation isadministered by intravitreal, subconjunctival, topical, or subtenonadministration.

Also described herein is a method for preventing an ocular disease in ahuman subject, the method comprising: (a) diluting a liquid formulationconsisting essentially of about 2% (w/w) rapamycin, about 4% (w/w)ethanol, and about 94% (w/w) PEG 400 with aqueous liquid to produce adiluted liquid formulation; and (b) administering to the human subjectby intraocular or periocular delivery a volume of the diluted liquidformulation, wherein an amount of rapamycin effective to prevent theocular disease is administered to the subject. In some variations, theformulation is a clear solution. In some variations, the dilution is upto about a 1.4-fold dilution. In some variations, the liquid formulationis diluted up to about 1.2 fold. In some variations, the aqueous liquidis saline. In some variations, the diluted liquid formulation isadministered by intravitreal, subconjunctival, topical, or subtenonadministration.

In some variations of each of the aforementioned methods and/orformulations, as well as other methods and/or formulations describedherein, the ocular disease is a disease of the retina. In somevariations, the ocular disease is wet age-related macular degeneration.In some variations, the ocular disease is dry age-related maculardegeneration. In some variations, the ocular disease is macular edema(including, without limitation, diabetic macular edema).

The present invention provides methods of treating macular edema in ahuman subject by repeated administration of rapamycin, the methodcomprising: administering two or more doses of a rapamycin formulationto an eye of the human subject, wherein the period between consecutivedoses is at least 8 weeks, and the cumulative amount of rapamycin in thetwo or more doses is effective to treat macular edema in the humansubject for an extended period of at least 16 weeks. In someembodiments, the period between consecutive doses is at least 12 weeks,and the cumulative amount of rapamycin in the two or more doses iseffective to treat macular edema in the human subject for an extendedperiod of at least 24 weeks. In some embodiments, the two or more dosesof the rapamycin formulation are administered by subconjunctivalplacement. In some embodiments, the rapamycin formulation is a solutionof rapamycin dissolved in a solvent system. In some embodiments, thesolvent system comprises polyethylene glycol. In some embodiments, thesolvent system further comprises ethanol. In some embodiments, therapamycin formulation comprises a suspension of particles of rapamycin.

The present invention also provides methods of treating diabetic macularedema in a human subject by repeated administration of rapamycin, themethod comprising: administering two or more doses of a rapamycinformulation to an eye of the human subject, wherein the rapamycinformulation is a solution comprising rapamycin and polyethylene glycol,the rapamycin formulation is administered by placement of each dosebetween a sclera and a conjunctiva of the eye of the subject, the periodbetween consecutive doses is at least 8 weeks, and wherein thecumulative amount of rapamycin in the two or more doses is effective totreat diabetic macular edema in the human subject for an extended periodof at least 16 weeks. In some embodiments, the period betweenconsecutive doses is at least 12 weeks and the cumulative amount ofrapamycin in the two or more doses is effective to treat diabeticmacular edema in the human subject for an extended period of at least 24weeks. In some embodiments, the rapamycin formulation further comprisesethanol. In a subset of these embodiments, each dose of the rapamycinformulation contains between about 200 μg and about 2,000 μg ofrapamycin, while in some preferred embodiments, each dose of therapamycin formulation contains about 220 μg, about 440 μg, about 880 μg,or about 1320 μg of rapamycin. In some preferred embodiments, therapamycin formulation comprises: (a) about 2% (w/w) rapamycin, about 4%(w/w) ethanol, and about 94% (w/w) polyethylene glycol 400 (PEG 400);(b) about 4% (w/w) rapamycin, about 4% (w/w) ethanol, and about 92%(w/w) polyethylene glycol 400 (PEG 400); or (c) about 1% (w/w)rapamycin, about 4% (w/w) ethanol, and about 95% (w/w) polyethyleneglycol 400 (PEG 400). In some embodiments, the rapamycin formulationconsists essentially of: (a) about 2% (w/w) rapamycin, about 4% (w/w)ethanol, and about 94% (w/w) polyethylene glycol 400 (PEG 400); (b)about 4% (w/w) rapamycin, about 4% (w/w) ethanol, and about 92% (w/w)polyethylene glycol 400 (PEG 400); or (c) about 1% (w/w) rapamycin,about 4% (w/w) ethanol, and about 95% (w/w) polyethylene glycol 400 (PEG400).

Moreover, the present invention provides liquid formulations comprising:(a) between about 0.001% (w/w) to about 1.0% (w/w) rapamycin; (b) afirst component selected from the group consisting of ethoxlyatedp-tert-octylphenol formaldehyde polymer, and polyoxyl 35 castor oil; and(c) a second component selected from the group consisting of glycerolcaprylate, and glycerol caprate. In some embodiments, the liquidformulations further comprise (d) a third component selected from thegroup consisting of ethanol, phosphatidylcholine in propyleneglycol/ethanol carrier, and propylene glycol monolaurate. In someembodiments, the liquid formulations further comprise water. In somepreferred embodiments, the liquid formulations comprise between about0.01% (w/w) to about 0.1% (w/w) rapamycin. In addition the presentinvention provides methods for treating an ocular disease in a humansubject, the method comprising administering to the human subject bytopical delivery to an eye a volume of the liquid formulation containingan amount of rapamycin effective to treat the ocular disease in thehuman subject. In some embodiments, the ocular disease is selected fromthe group consisting of age-related macular degeneration, macular edema,dry eye, allergic conjunctivitis, scleritis, uveitis, and cornealtransplant. In a subset of these embodiments, the age-related maculardegeneration is wet age-related macular degeneration. In someembodiments, the period of time between consecutive administrations ofthe liquid formulation is at least about 1 day, while in others theperiod of time between consecutive administrations of the liquidformulation is at least about 7 days or at least about 14 days.

In addition the present invention provides methods for treating anocular disease in a human subject, the method comprising administeringto the human subject by topical delivery a volume of a liquidformulation containing an amount of a therapeutic agent effective toprevent or treat the ocular disease in the human subject, wherein theliquid formulation when topically applied to a rabbit eye in a volume ofat least about 40 μl a day for six days delivers an amount of thetherapeutic agent sufficient to achieve one or more of: (a) an averageconcentration of the therapeutic agent in the cornea of the rabbit eyeat day 8 equivalent to a rapamycin concentration of at least 0.01 ng/g;(b) an average concentration of the therapeutic agent in the retinachoroid of the rabbit eye at day 8 equivalent to a rapamycinconcentration of at least 0.01 ng/g; and (c) an average concentration ofthe therapeutic agent in the sclera of the rabbit eye at day 8equivalent to a rapamycin concentration of at least 0.01 ng/g. In someembodiments, the liquid formulation when topically applied achieves oneor more of (a), (b), and (c), and further achieves: (d) an averageconcentration of therapeutic agent in the blood of the rabbit at day 8less than the average concentration of the therapeutic agent in one ormore of the cornea, the retina choroid and the sclera. In someembodiments, the therapeutic agent is rapamycin, or a pharmaceuticallyacceptable salt or ester thereof, while in others the therapeutic agentis dasatinib, or a pharmaceutically acceptable salt or ester thereof. Insome preferred embodiments, the ocular disease is selected from thegroup consisting of age-related macular degeneration, macular edema, dryeye, allergic conjunctivitis, scleritis, uveitis, and cornealtransplant. In some preferred embodiments, the age-related maculardegeneration is wet age-related macular degeneration. In someembodiments, the period of time between consecutive administrations ofthe liquid formulation is at least about 1 day, while in others theperiod of time between consecutive administrations of the liquidformulation is at least about 7 days, or at least about 14 days. In asubset of these embodiments, the liquid formulation consists essentiallyof (a) about 2% (w/w) rapamycin, about 4% (w/w) ethanol, and about 94%(w/w) PEG 400; (b) about 4% (w/w) rapamycin, about 4% (w/w) ethanol, andabout 92% (w/w) PEG 400; or (c) about 1% (w/w) rapamycin, about 4% (w/w)ethanol, and about 95% (w/w) PEG 400.

The present invention also provides methods for treating an oculardisease in a human subject, the method comprising topicallyadministering to the subject a liquid formulation comprising atherapeutic agent, wherein the liquid formulation, when a volume of atleast about 40 μL is applied topically to a rabbit eye, delivers anamount of the therapeutic agent sufficient to achieve one or more of:(a) an average concentration of the therapeutic agent in the cornea ofthe rabbit eye equivalent to an average rapamycin concentration of atleast 0.01 ng/g for at least about seven days after administration ofthe liquid formulation; (b) an average concentration of therapeuticagent in the retina choroid of the rabbit eye equivalent to an averagerapamycin concentration of at least 0.01 ng/g for at least about sevendays after administration of the liquid formulation; and (c) an averageconcentration of the therapeutic agent in the sclera of the rabbit eyeequivalent to an average rapamycin concentration of at least 0.01 ng/gfor at least about seven days after administration of the liquidformulation. In some embodiments, the liquid formulation when topicallyapplied achieves one or more of (a), (b), and (c), and further achieves:(d) an average concentration of therapeutic agent in the blood of therabbit at day seven less than the average concentration of thetherapeutic agent in one or more of the cornea, the retina choroid andthe sclera. In some embodiments, the therapeutic agent is rapamycin or apharmaceutically acceptable salt or ester thereof. In some preferredembodiments, the ocular disease is selected from the group consisting ofage-related macular degeneration, macular edema, dry eye, allergicconjunctivitis, scleritis, uveitis, and corneal transplant. In someparticularly preferred embodiments, the age-related macular degenerationis wet age-related macular degeneration. In some embodiments, the liquidformulation comprises between about 0.001% (w/w) to about 10% (w/w)rapamycin. In a subset of these embodiments, the liquid formulationcomprises between about 0.01% (w/w) to about 1.0% (w/w) rapamycin. Insome embodiments, the liquid formulation further comprises polyethyleneglycol. In some preferred embodiments, the period of time betweenconsecutive administrations of the liquid formulation is at least about1 day, while in others the period of time between consecutiveadministrations of the liquid formulation is at least about 7 days or atleast about 14 days.

Moreover the present invention provides methods of administering by aperiocular route a liquid formulation to an eye of a subject withoutcausing chemosis of the eye, comprising: administering less than 40 μlof the liquid formulation between a sclera and a conjunctiva of the eyeof the subject, wherein the liquid formulation comprises a therapeuticagent and a solvent, and wherein the liquid formulation has ahygroscopicity that is 80% or greater than the hygroscopicity of aliquid formulation consisting essentially of about 2% (w/w) rapamycin,about 4% (w/w) ethanol, and about 94% (w/w) polyethylene glycol 400 (PEG400). As used herein, the term “chemosis” refers to edema of theconjunctiva of the eye, forming a swelling partly or fully around theiris of the eye. In some embodiments, the therapeutic agent israpamycin. In some embodiments, the solvent comprises polyethyleneglycol, while in a subset of these embodiments, the polyethylene glycolis polyethylene glycol 400 (PEG 400). In some embodiments, about 1 μg toabout 5,000 μg rapamycin is administered to the subject. In a subset ofthese embodiments, about 220 μg, about 440 μg, about 880 μg, or about1320 μg rapamycin is administered to the subject. Also provided aremethods for treating an ocular disease in a human subject, wherein theocular disease is selected from the group consisting of age-relatedmacular degeneration, macular edema and dry eye, the method comprisingadministering to the subject, an amount of the therapeutic agenteffective to treat the ocular disease in the human subject. In someembodiments, the ocular disease is age-related macular degeneration,which in preferred embodiments is wet age-related macular degeneration.In some embodiments, the ocular disease is macular edema, which inpreferred embodiments is diabetic macular edema. In other preferredembodiments, the ocular disease is dry eye.

In addition, the present invention provides methods for treating anocular disease in a human subject, the method comprising: (a)administering to the human subject a volume of a first formulation byintraocular or periocular delivery to the subject, wherein the volume ofthe first formulation comprises an amount of ranibizumab or bevacizumab;and (b) administering to the human subject a volume of a secondformulation by periocular delivery to the subject, wherein the volume ofthe second formulation comprises an amount of rapamycin, or apharmaceutically acceptable salt or ester thereof; wherein the amount ofranibizumab or bevacizumab and the amount of rapamycin or apharmaceutically acceptable salt or ester thereof, together areeffective to treat the ocular disease when administered to the humansubject. In some embodiments, the first formulation is administered byintravitreal placement, and the second formulation is administered bysubconjunctival placement. In some embodiments, step (a) is coincidentwith step (b) (same doctor visit or occurring on the same day). In otherembodiments, step (b) is subsequent to step (a) (different doctor visitsor occurring on different days). In some embodiments, the ocular diseaseis age-related macular degeneration, or macular edema. In some preferredembodiments, the age-related macular degeneration is wet age-relatedmacular degeneration, or the macular edema is diabetic macular edema. Insome embodiments, the volume of the first formulation contains betweenabout 100 μg and about 500 μg of ranibizumab or bevacizumab. In someembodiments, the volume of the second formulation contains between about200 μg and about 2000 μg of rapamycin.

The present invention also provides methods for treating an oculardisease in a human subject, the method comprising: (a) administering tothe human subject by intraocular or periocular delivery a volume of afirst formulation comprising an amount of a first therapeutic agent; and(b) administering to the human subject by periocular delivery a volumeof a second formulation comprising an amount of rapamycin, wherein thesecond formulation consists essentially of rapamycin, ethanol, andpolyethylene glycol 400 (PEG 400), and wherein the amount of the firsttherapeutic agent and the amount of rapamycin together are effective totreat the ocular disease when administered to the human subject. In someembodiments, wherein the first therapeutic agent is selected from thegroup consisting of ranibizumab, bevacizumab, and verteporfin. In someembodiments, the second formulation consists essentially of: (a) about2% (w/w) rapamycin, about 4% (w/w) ethanol, and about 94% (w/w) PEG 400;(b) about 4% (w/w) rapamycin, about 4% (w/w) ethanol, and about 92%(w/w) PEG 400; or (c) about 1% (w/w) rapamycin, about 4% (w/w) ethanol,and about 95% (w/w) PEG 400. In some embodiments, the administration ofthe first formulation is by a route selected from the group consistingof intravitreal, subconjunctival, and subtenon placement, and theadministration of the second formulation is by subconjunctival orsubtenon placement. In some embodiments, step (a) is coincident withstep (b) (same doctor visit or occurring on the same day). In otherembodiments, step (b) is subsequent to step (a) (different doctor visitsor occurring on different days). In some embodiments, the ocular diseaseis age-related macular degeneration or macular edema. In some preferredembodiments, the age-related macular degeneration is wet age-relatedmacular degeneration. In some embodiments, the ocular disease is macularedema. In some preferred embodiments, the macular edema is diabeticmacular edema.

Furthermore the present invention provides methods for treating wetage-related macular degeneration or diabetic macular edema in a humansubject, the method comprising: (a) administering to the human subject avolume of a first formulation by intravitreal placement, wherein thevolume of the first formulation comprises an amount of ranibizumab orbevacizumab; and (b) administering to the human subject a volume of asecond formulation by subconjunctival placement, wherein the volume ofthe second formulation comprises an amount of rapamycin, or apharmaceutically acceptable salt or ester thereof; wherein the amount ofranibizumab or bevacizumab and the amount of rapamycin or apharmaceutically acceptable salt or ester thereof, together areeffective to treat wet age-related macular degeneration or diabeticmacular edema when administered to the human subject. In someembodiments, step (a) is coincident with step (b) (same doctor visit oroccurring on the same day). In other embodiments, step (b) is subsequentto step (a) (different doctor visits or occurring on different days). Insome embodiments, the volume of the first formulation contains betweenabout 100 μg and about 500 μg of ranibizumab or bevacizumab. In someembodiments, the volume of the second formulation contains between about200 μg and about 2000 μg of rapamycin. In some embodiments, the secondformulation consists essentially of (a) about 2% (w/w) rapamycin, about4% (w/w) ethanol, and about 94% (w/w) polyethylene glycol 400 (PEG 400);(b) about 4% (w/w) rapamycin, about 4% (w/w) ethanol, and about 92%(w/w) polyethylene glycol 400 (PEG 400); or (c) about 1% (w/w)rapamycin, about 4% (w/w) ethanol, and about 95% (w/w) polyethyleneglycol 400 (PEG 400).

Moreover the present invention provides liquid formulations comprising(i) a solvent selected from the group consisting of N-methylpyrrolidone(NMP), dimethyl acetamine (DMA), and dimethyl sulfoxide (DMSO); (ii)poly(D,L-lactide-co-glycolide) (PLGA), or a solvent selected from thegroup consisting of propylene glycol (PG), polyethylene glycol 600 (PEG600), polyethylene glycol 400 (PEG 400); and (iii) a therapeutic agent.In some embodiments, the liquid formulation when: (a) injected betweenthe sclera and conjunctiva of a rabbit eye delivers an amount of thetherapeutic agent sufficient to achieve, for a period of time of atleast 30 days following administration of the liquid formulation, one orboth of the following: (i) an average concentration of therapeutic agentin the vitreous of the rabbit eye equivalent to a rapamycinconcentration of at least 0.01 ng/ml; or (ii) an average concentrationof therapeutic agent in the retina choroid tissues of the rabbit eyeequivalent to a rapamycin concentration of at least 0.01 ng/g; (b)administered by subtenon injection to a rabbit eye delivers an amount ofthe therapeutic agent sufficient to achieve, for a period of time of atleast 30 days following administration of the liquid formulation, one orboth of the following: (i) an average concentration of therapeutic agentin the vitreous of the rabbit eye equivalent to a rapamycinconcentration of at least 0.01 ng/ml; or (ii) an average concentrationof therapeutic agent in the retina choroid tissues of the rabbit eyeequivalent to a rapamycin concentration of at least 0.01 ng/g; or (c)injected into the vitreous of a rabbit eye delivers an amount of thetherapeutic agent sufficient to achieve, for a period of time of atleast 30 days following administration of the liquid formulation, one orboth of the following: (a) an average concentration of therapeutic agentin the vitreous of the rabbit eye equivalent to a rapamycinconcentration of at least 100 ng/ml; or (b) an average concentration oftherapeutic agent in the retina choroid tissues of the rabbit eyeequivalent to a rapamycin concentration of at least 0.01 ng/g. In someembodiments, the liquid formulation comprises between about 0.001% (w/w)to about 10% (w/w) of the therapeutic agent. In some embodiments, thetherapeutic agent is rapamycin, or a pharmaceutically acceptable salt orester thereof. In other embodiments, the therapeutic agent is dasatinib,or a pharmaceutically acceptable salt or ester thereof. In someembodiments, the liquid formulation consists essentially of (a) about 4%(w/w) rapamycin, about 4% (w/w) ethanol, and about 92% (w/w) PEG 400; or(b) about 1.0-4.0% (w/w) rapamycin, about 2.0-4.0% (w/w) ethanol, about20-30% (w/w) normal saline, and quantity of PEG 400 sufficient for thetotal formula weight percentage to equal 100%. The present inventionfurther provides methods for treating an ocular disease in a humansubject, the method comprising administering to the human subject byintraocular or periocular delivery a volume of the liquid formulationcontaining an amount of the therapeutic agent effective to treat theocular disease in the human subject. In some embodiments, the oculardisease is age-related macular degeneration, macular edema, or dry eye.In some preferred embodiments, the age-related macular degeneration iswet age-related macular degeneration. In other preferred embodiments,the macular edema is diabetic macular edema. In still further preferredembodiments, the ocular disease is dry eye. In some particularlypreferred embodiments, the administration of the liquid formulation isby intravitreal, subconjunctival, or subtenon administration.

Additionally, the present invention provides methods for treating anocular disease in a human subject, the method comprising administeringto the human subject a volume of a liquid formulation comprising betweenabout 200 μg and about 5000 μg of rapamycin, or a pharmaceuticallyacceptable salt or ester thereof. In some embodiments, the volume of theliquid formulation contains between about 200 μg and about 1500 μg ofrapamycin. In some embodiments, when administration is by placement ofthe liquid formulation in the vitreous of the subject, the volume of theliquid formulation is about 100 μl or less, preferably about 25 μl orless. In some embodiments, when administration is by placement of theliquid formulation between the sclera and conjunctiva of the subject,the volume of the liquid formulation is about 1,000 μl or less,preferably about 50 μl or less. In some embodiments, when administrationis by placement of the liquid formulation in the subtenon space of thesubject, the volume of the liquid formulation is about 5,000 μl or less,preferably about 100 μl or less. In some embodiments, the ocular diseaseis age-related macular degeneration, macular edema, or dry eye. In somepreferred embodiments, the age-related macular degeneration is wetage-related macular degeneration. In some preferred embodiments, themacular edema is diabetic macular edema.

The present invention also provides a solid formulation comprisingrapamycin, polyvinylpyrrolidone, a phospholipid solution, andpolyethylene glycol. In some embodiments, the formulation furthercomprises one or both of gamma tocopherol and ascorbyl palmatate. Infurther embodiments, methods are provided for treating an ocular diseasein a human subject, the method comprising administering to an eye of thehuman subject by topical or periocular delivery a volume of the solidformulation containing an amount of rapamycin effective to treat theocular disease in the human subject. In some embodiments, the oculardisease is age-related macular degeneration, macular edema, or dry eye.In some embodiments, the amount of rapamycin in the solid formulation iseffective to treat the ocular disease in the human subject for anextended period of at least two weeks, at least four weeks, at least sixweeks or at least eight weeks.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts the average level of rapamycin in the retina choroidtissue (ng/g) of rabbit eyes at 14, 30, 60, and 90 days aftersubconjunctival injection of 40 μl of 2% solution of rapamycin inethanol and PEG 400, 1.47% solution of rapamycin in ethanol and PEG 400,or 1.38% solution of rapamycin in ethanol and PEG 400 or 20 μl of a 4%solution of rapamycin in ethanol and PEG 400.

FIG. 2 depicts the level of rapamycin in the retina choroid tissue(ng/g) of rabbit eyes at 3, 14, 30, 45, and 60 days after intravitrealinjection of 6 μl of 2% solution of rapamycin in ethanol and PEG 400 or8.5 μl of 1.47% solution of rapamycin in ethanol and PEG 400.

FIGS. 3A-3D depict the concentration of rapamycin present in (A) wholeblood (ng/ml), (B) sclera (ng/g), (C) retina choroid (ng/g), and (D)vitreous fluid (ng/mL) at 3, 7, 14, 30, 60, and 90 days after eithersubconjunctival or subtenous injection of 30 μl of 2% solution ofrapamycin in ethanol and PEG 400.

FIG. 4 depicts the concentration of rapamycin present in tissue or fluidof rabbit eyes after topical administration of one eye drop (about 40μL) of a 2% rapamycin solution.

FIG. 5 depicts the concentration of rapamycin present in tissue or fluidof rabbit eyes after topical administration by eye drops of a 0.07%rapamycin solution.

FIG. 6 depicts the concentration of rapamycin present in eye tissue(ng/g) of rabbit eyes after topical administration by eye drops of a0.09% rapamycin solution.

FIG. 7 depicts the concentration of rapamycin present in eye tissue orfluid (ng/g or ng/ml) after topical administration by eye drops of a0.08% rapamycin solution comprising phosal, capmul, tyloxapol and water.

FIG. 8 depicts the concentration of rapamycin present in eye tissue orfluid (ng/g or ng/ml) after topical administration by eye drops of a0.17% rapamycin solution comprising cremophor, capmul, ethanol, andwater.

FIG. 9 depicts the concentration of rapamycin present in eye tissue orblood (ng/g or ng/ml) after topical administration by eye drops of anon-aqueous solution comprising 0.5%, 1.0% or 4.0% rapamycin.

FIGS. 10A-B depict the concentration of rapamycin present in eye tissueor blood (ng/g or ng/ml) after topical administration by eye drops of anon-aqueous solution or an aqueous solution comprising about 0.2%rapamycin.

FIG. 11 depicts the concentration of rapamycin present in eye tissue orfluid (ng/g or ng/ml) after topical administration by eye drops of anon-aqueous solution comprising about 0.2% rapamycin.

FIG. 12 depicts the concentration of rapamycin present in eye tissue orfluid (ng/g or ng/ml) after intravitreal administration of liquidformulation comprising about 0.6% rapamycin.

FIG. 13 depicts the concentration of rapamycin present in eye tissue orfluid (ng/g or ng/ml) after periocular administration of solidformulation comprising about 10% rapamycin.

FIG. 14 depicts the concentration of dasatanib present in eye tissue orfluid (ng/g or ng/ml) after intravitreal administration of liquidformulation comprising about 5% dasatanib.

DETAILED DESCRIPTION

Described herein are compositions, liquid formulations and methodsrelating to delivery of therapeutic agents to a subject, including butnot limited to a human subject or to the eye of a subject. Thesecompositions, liquid formulations, and methods may be used for thetreatment, prevention, inhibition, delaying onset of, or causingregression of diseases and conditions of the eye including but notlimited to diseases or conditions of the posterior segment, includingbut not limited to choroidal neovascularization; macular degeneration;age-related macular degeneration, including wet AMD and dry AMD; retinalangiogenesis; chronic uveitis; and other retinoproliferative conditions.In some variations, the compositions, liquid formulations, and methodsare used for the treatment of the aforementioned diseases or conditionsof the eye.

Herein are described (1) the therapeutic agents that may be delivered toa subject, including but not limited to a human subject or an eye of asubject using the compositions, liquid formulations, and methodsdescribed herein, (2) the diseases and conditions that may be treated,prevented, inhibited, onset delayed, or regression caused by delivery ofthe therapeutic agents, (3) liquid formulations that may be used todeliver the therapeutic agents, (4) routes of administration fordelivery of the liquid formulations, (5) extended delivery oftherapeutic agents including but not limited to rapamycin, and (6)description of the treatment of CNV and wet AMD by delivery of rapamycinto a subject, including but not limited to a human subject or to the eyeof a subject for an extended period of time using the describedcompositions and liquid formulations.

The term “about,” as used herein, refers to the level of accuracy thatis obtained when the methods described herein, such as the methods inthe examples, are used. However, by “about” a certain amount of acomponent of a formulation is meant 90-110% of the amount stated.

Therapeutic Agents

Most generally, any compounds and compositions currently known or yet tobe discovered that are useful in treating, preventing, inhibiting,delaying the onset of, or causing the regression of the diseases andconditions described herein may be therapeutic agents for use in thecompositions, liquid formulations, and methods described herein.

Therapeutic agents that may be used include compounds that act bybinding members of the immunophilin family of cellular proteins. Suchcompounds are known as “immunophilin binding compounds.” Immunophilinbinding compounds include but are not limited to the “limus” family ofcompounds. Examples of limus compounds that may be used include but arenot limited to cyclophilins and FK506-binding proteins (FKBPs),including sirolimus (rapamycin) and its water soluble analog SDZ-RAD(Novartis), TAFA-93 (Isotechnika), tacrolimus, everolimus, RAD-001(Novartis), pimecrolimus, temsirolimus, CCI-779 (Wyeth), AP23841(Ariad), AP23573 (Ariad), and ABT-578 (Abbott Laboratories). Limuscompound analogs and derivatives that may be used include but are notlimited to the compounds described in U.S. Pat. Nos. 5,527,907;6,376,517; and 6,329,386 and U.S. patent application Ser. No.09/950,307, each of which is incorporated herein by reference in theirentirety. Therapeutic agents also include analogs, prodrugs, salts andesters of limus compounds.

The terms rapamycin, rapa, and sirolimus are used interchangeablyherein.

Other rapamycin derivatives that may be used include, withoutlimitation, 7-epi-rapamycin, 7-thiomethyl-rapamycin,7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin,7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-desmethyl-rapamycin,mono- and di-ester derivatives of rapamycin, 27-oximes of rapamycin;42-oxo analog of rapamycin; bicyclic rapamycins; rapamycin dimers; silylethers of rapamycin; rapamycin arylsulfonates and sulfamates,mono-esters and di-esters at positions 31 and 42, 30-demethoxyrapamycin, and other derivatives described in Vezina et al., “Rapamycin(AY-22,989), A New Antifungal Antibiotic. I. Taxonomy Of The ProducingStreptomycete And Isolation Of The Active Principle” J. Antibiot.(Tokyo) 28:721-726 (1975); Sehgal et al., “Rapamycin (AY-22,989), A NewAntifungal Antibiotic. II. Fermentation, Isolation And Characterization”J. Antibiot. (Tokyo) 28:727-732 (1975); Sehgal et al.,“Demethoxyrapamycin (AY-24,668), A New Antifungal Antibiotic” J.Antibiot. (Tokyo) 36:351-354 (1983); and Paiva et al., “Incorporation OfAcetate, Propionate, And Methionine Into Rapamycin By Streptomyceteshygroscopicus” J Nat Prod 54:167-177 (1991), WO 92/05179, EP 467606,Caufield et al., “Hydrogenated Rapamycin Derivatives” U.S. Pat. No.5,023,262; Kao et al., “Bicyclic Rapamycins” U.S. Pat. No. 5,120,725;Kao et al., “Rapamycin Dimers” U.S. Pat. No. 5,120,727; Failli et al.,“Silyl Ethers Of Rapamycin” U.S. Pat. No. 5,120,842; Failli et al.,“Rapamycin 42-Sulfonates And 42-(N-carboalkoxy) Sulfamates Useful AsImmunosuppressive Agents” U.S. Pat. No. 5,177,203; Nicolaou et al.,“Total Synthesis Of Rapamycin” J. Am. Chem. Soc. 115: 4419-4420 (1993);Romo et al, “Total Synthesis Of (−) Rapamycin Using An Evans-TishchenkoFragment Coupling” J. Am. Chem. Soc. 115:7906-7907 (1993); and Haywardet al, “Total Synthesis Of Rapamycin Via A Novel Titanium-Mediated AldolMacrocyclization Reaction” J. Am. Chem. Soc., 115:9345-9346 (1993), eachof which is incorporated herein by reference in its entirety.

The limus family of compounds may be used in the compositions, liquidformulations and methods for the treatment, prevention, inhibition,delaying the onset of, or causing the regression ofangiogenesis-mediated diseases and conditions of the eye, includingchoroidal neovascularization. The limus family of compounds may be usedto prevent, treat, inhibit, delay the onset of, or cause regression ofAMD, including wet AMD. Rapamycin and rapamycin derivatives and analogsmay be used to prevent, treat, inhibit, delay the onset of, or causeregression of angiogenesis-mediated diseases and conditions of the eye,including choroidal neovascularization. Rapamycin may be used toprevent, treat, inhibit, delay the onset of, or cause regression of AMD,including wet AMD. In some variations, a member of the limus family ofcompounds or rapamycin is used to treat wet AMD or angiogenesis-mediateddiseases and conditions of the eye including choroidalneovascularization.

Other therapeutic agents that may be used include those disclosed in thefollowing patents and publications, the contents of each of which isincorporated herein by reference in its entirety: PCT publication WO2004/027027, published Apr. 1, 2004, titled Method of inhibitingchoroidal neovascularization, assigned to Trustees of the University ofPennsylvania; U.S. Pat. No. 5,387,589, issued Feb. 7, 1995, titledMethod of Treating Ocular Inflammation, with inventor Prassad Kulkami,assigned to University of Louisville Research Foundation; U.S. Pat. No.6,376,517, issued Apr. 23, 2003, titled Pipecolic acid derivatives forvision and memory disorders, assigned to GPI NIL Holdings, Inc; PCTpublication WO 2004/028477, published Apr. 8, 2004, titled Methodsubretinal administration of therapeutics including steroids: method forlocalizing pharmadynamic action at the choroid and retina; and relatedmethods for treatment and or prevention of retinal diseases, assigned toInnorx, Inc; U.S. Pat. No. 6,416,777, issued Jul. 9, 2002, titledOphthalmic drug delivery device, assigned to Alcon Universal Ltd; U.S.Pat. No. 6,713,081, issued Mar. 30, 2004, titled Ocular therapeuticagent delivery device and methods for making and using such devices,assigned to Department of Health and Human Services; U.S. Pat. No.5,100,899, issued Mar. 31, 1992, titled Methods of inhibiting transplantrejection in mammals using rapamycin and derivatives and prodrugsthereof.

Other therapeutic agents that may be used include pyrrolidine,dithiocarbamate (NFκB inhibitor); squalamine; TPN 470 analogue andfumagillin; PKC (protein kinase C) inhibitors; Tie-1 and Tie-2 kinaseinhibitors; inhibitors of VEGF receptor kinase; proteosome inhibitorssuch as Velcade™ (bortezomib, for injection; ranibuzumab (Lucentis™) andother antibodies directed to the same target; pegaptanib (Macugen™);vitronectin receptor antagonists, such as cyclic peptide antagonists ofvitronectin receptor-type integrins; α-v/β-3 integrin antagonists;α-v/β-1 integrin antagonists; thiazolidinediones such as rosiglitazoneor troglitazone; interferon, including γ-interferon or interferontargeted to CNV by use of dextran and metal coordination; pigmentepithelium derived factor (PEDF); endostatin; angiostatin; tumistatin;canstatin; anecortave acetate; acetonide; triamcinolone;tetrathiomolybdate; RNA silencing or RNA interference (RNAi) ofangiogenic factors, including ribozymes that target VEGF expression;Accutane™ (13-cis retinoic acid); ACE inhibitors, including but notlimited to quinopril, captopril, and perindozril; inhibitors of mTOR(mammalian target of rapamycin); 3-aminothalidomide; pentoxifylline;2-methoxyestradiol; colchicines; AMG-1470; cyclooxygenase inhibitorssuch as nepafenac, rofecoxib, diclofenac, rofecoxib, NS398, celecoxib,vioxx, and (E)-2-alkyl-2(4-methanesulfonylphenyl)-1-phenylethene; t-RNAsynthase modulator; metalloprotease 13 inhibitor; acetylcholinesteraseinhibitor; potassium channel blockers; endorepellin; purine analog of6-thioguanine; cyclic peroxide ANO-2; (recombinant) arginine deiminase;epigallocatechin-3-gallate; cerivastatin; analogues of suramin; VEGFtrap molecules; apoptosis inhibiting agents; Visudyne™, snET2 and otherphoto sensitizers, which may be used with photodynamic therapy (PDT);inhibitors of hepatocyte growth factor (antibodies to the growth factoror its receptors, small molecular inhibitors of the c-met tyrosinekinase, truncated versions of HGF e.g. NK4).

In some variations the therapeutic agent is a BCR/ABL inhibitor. In somevariations the therapeutic agent is a Src family tyrosine kinasesinhibitor. In some variations the therapeutic agent is a dual BCR/ABLand Src family tyrosine kinases inhibitor. In some variations the dualBCR/ABL and Src family tyrosine kinases inhibitor is dasatinib (i.e.,BMS-354825, Sprycel®, orN-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)-1-piperazinyl]-2-methyl-4-pyrimidinyl]amino]-5-thiazolecarboxamide monohydrate) or a derivative, an analog, or apharmaceutically acceptable salt or ester thereof. In some variations,dasatinib is used to treat, prevent, inhibit, or delay onset orregression of an ocular disease. In some variations, the ocular diseaseis age-related macular degeneration (AMD). In some variations, the AMDis wet AMD. In some variations, the AMD is dry AMD. In some variations,the ocular disease is macular edema including without limitationdiabetic macular edema.

Other therapeutic agents that may be used include anti-inflammatoryagents, including, but not limited to nonsteroidal anti-inflammatoryagents and steroidal anti-inflammatory agents. In some variations,active agents that may be used in the liquid formulations areace-inhibitors, endogenous cytokines, agents that influence basementmembrane, agents that influence the growth of endothelial cells,adrenergic agonists or blockers, cholinergic agonists or blockers,aldose reductase inhibitors, analgesics, anesthetics, antiallergics,antibacterials, antihypertensives, pressors, antiprotozoal agents,antiviral agents, antifungal agents, anti-infective agents, antitumoragents, antimetabolites, and antiangiogenic agents.

Steroidal therapeutic agents that may be used include but are notlimited to 21-acetoxypregnenolone, alclometasone, algestone, amcinonide,beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol,clobetasone, clocortolone, cloprednol, corticosterone, cortisone,cortivazol, deflazacort, desonide, desoximetasone, dexamethasone,diflorasone, diflucortolone, difluprednate, enoxolone, fluazacort,flucloronide, flumethasone, flunisolide, fluocinolone acetonide,fluocinonide, fluocortin butyl, fluocortolone, fluorometholone,fluperolone acetate, fluprednidene acetate, fluprednisolone,flurandrenolide, fluticasone propionate, formocortal, halcinonide,halobetasol propionate, halometasone, halopredone acetate,hydrocortamate, hydrocortisone, loteprednol etabonate, mazipredone,medrysone, meprednisone, methylprednisolone, mometasone furoate,paramethasone, prednicarbate, prednisolone, prednisolone25-diethylamino-acetate, prednisolone sodium phosphate, prednisone,prednival, prednylidene, rimexolone, tixocortol, triamcinolone,triamcinolone acetonide, triamcinolone benetonide, triamcinolonehexacetonide, and any of their derivatives.

In some variations, cortisone, dexamethasone, fluocinolone,hydrocortisone, methylprednisolone, prednisolone, prednisone, andtriamcinolone, or their derivatives, may be used. The liquid formulationmay include a combination of two or more steroidal therapeutic agents.

In one nonlimiting example, the steroidal therapeutic agents mayconstitute from about 0.05% to about 50% by weight of the liquidformulation. In another nonlimiting example, the steroid constitutesfrom about 0.05% to about 10%, between about 10% to about 20%; betweenabout 30% to about 40%; or between about 40% to about 50% by weight ofthe liquid formulation.

Other nonlimiting examples of therapeutic agents that may be usedinclude but are not limited to anaesthetics, analgesics, celltransport/mobility impending agents such as colchicines, vincristine,cytochalasin B and related compounds; carbonic anhydrase inhibitors suchas acetazolamide, methazolamide, dichlorphenamide, diamox andneuroprotectants such as nimodipine and related compounds; antibioticssuch as tetracycline, chlortetracycline, bacitracin, neomycin,polymyxin, gramicidin, cephalexin, oxytetracycline, chloramphenicol,rifampicin, ciprofloxacin, aminosides, gentamycin, erythromycin andpenicillin, quinolone, ceftazidime, vancomycine imipeneme; antifungalssuch as amphotericin B, fluconazole, ketoconazole and miconazole;antibacterials such as sulfonamides, sulfadiazine, sulfacetamide,sulfamethizole and sulfisoxazole, nitrofurazone and sodium propionate;antivirals, such as idoxuridine, trifluorothymidine, trifluorouridine,acyclovir, ganciclovir, cidofovir, interferon, DDI, AZT, foscamet,vidarabine, irbavirin, protease inhibitors and anti-cytomegalovirusagents; antiallergenics such as sodium cromoglycate, antazoline,methapyriline, chlorpheniramine, cetirizine, pyrilamine andprophenpyridamine; synthetic gluocorticoids and mineralocorticoids andmore generally hormones forms derivating from the cholesterol metabolism(DHEA, progesterone, estrogens); non-steroidal anti-inflammatories suchas salicylate, indomethacin, ibuprofen, diclofenac, flurbiprofen,piroxicam and COX2 inhibitors; antineoplastics such as carmustine,cisplatin, fluorouracil; adriamycin, asparaginase, azacitidine,azathioprine, bleomycin, busulfan, carboplatin, carmustine,chlorambucil, cyclophosphamide, cyclosporine, cytarabine, dacarbazine,dactinomycin, daunorubicin, doxorubicin, estramustine, etoposide,etretinate, filgrastin, floxuridine, fludarabine, fluorouracil,florxymesterone, flutamide, goserelin, hydroxyurea, ifosfamide,leuprolide, levamisole, limustine, nitrogen mustard, melphalan,mercaptopurine, methotrexate, mitomycin, mitotane, pentostatin,pipobroman, plicamycin, procarbazine, sargramostin, streptozocin,tamoxifen, taxol, teniposide, thioguanine, uracil mustard, vinblastine,vincristine and vindesine; immunological drugs such as vaccines andimmune stimulants; insulin, calcitonin, parathyroid hormone and peptideand vasopressin hypothalamus releasing factor; beta adrenergic blockerssuch as timolol, levobunolol and betaxolol; cytokines, interleukines andgrowth factors epidermal growth factor, fibroblast growth factor,platelet derived growth factor, transforming growth factor beta, ciliaryneurotrophic growth factor, glial derived neurotrophic factor, NGF, EPO,PLGF, brain nerve growth factor (BNGF), vascular endothelial growthfactor (VEGF) and monoclonal antibodies or fragments thereof directedagainst such growth factors; anti-inflammatories such as hydrocortisone,dexamethasone, fluocinolone, prednisone, prednisolone,methylprednisolone, fluorometholone, betamethasone and triamcinolone;decongestants such as phenylephrine, naphazoline and tetrahydrazoline;miotics and anti-cholinesterases such as pilocarpine, carbachol,di-isopropyl fluorophosphate, phospholine iodine and demecarium bromide;mydriatics such as atropine sulphate, cyclopentolate, homatropine,scopolamine, tropicamide, eucatropine; sympathomimetics such asepinephrine and vasoconstrictors and vasodilators, anticlotting agentssuch as heparin, antifibrinogen, fibrinolysin, anticlotting activase,antidiabetic agents include acetohexamide, chlorpropamide, glipizide,glyburide, tolazamide, tolbutamide, insulin and aldose reductaseinhibitors, hormones, peptides, nucleic acids, saccharides, lipids,glycolipids, glycoproteins and other macromolecules include endocrinehormones such as pituitary, insulin, insulin-related growth factor,thyroid, growth hormones; heat shock proteins; immunological responsemodifiers such as muramyl dipeptide, cyclosporins, interferons(including alpha-, beta- and gamma-interferons), interleukin-2,cytokines, FK506 (an epoxy-pyrido-oxaazcyclotricosine-tetrone, alsoknown as Tacrolimus), tumor necrosis factor, pentostatin, thymopentin,transforming factor beta2, erythropoetin; antineogenesis proteins (e.g.anti VEGF, interferons), antibodies (monoclonal, polyclonal, humanized,etc.) or antibodies fragments, oligoaptamers, aptamers and genefragments (oligonucleotides, plasmids, ribozymes, small interference RNA(SiRNA), nucleic acid fragments, peptides), immunomodulators such asendoxan, thalidomide, tamoxifene; antithrombolytic and vasodilatoragents such as rtPA, urokinase, plasmin; nitric oxide donors, nucleicacids, dexamethasone, cyclosporin A, azathioprine, brequinar,gusperimus, 6-mercaptopurine, mizoribine, rapamycin, tacrolimus(FK-506), folic acid analogs (e.g., denopterin, edatrexate,methotrexate, piritrexim, pteropterin, Tomudex®, trimetrexate), purineanalogs (e.g., cladribine, fludarabine, 6-mercaptopurine, thiamiprine,thiaguanine), pyrimidine analogs (e.g., ancitabine, azacitidine,6-azauridine, carmofur, cytarabine, doxifluridine, emitefur,enocitabine, floxuridine, fluorouracil, gemcitabine, tegafur)fluocinolone, triaminolone, anecortave acetate, fluorometholone,medrysone, and prednislone. In some variations the immunosuppressiveagent is dexamethasone. In some variations the immunosuppressive agentis cyclosporin A.

In some variations the formulation comprises a combination of one ormore therapeutic agents.

Other nonlimiting examples of therapeutic agents that may be used in theformulations described herein include antibacterial antibiotics,aminoglycosides (e.g., amikacin, apramycin, arbekacin, bambermycins,butirosin, dibekacin, dihydrostreptomycin, fortimicin(s), gentamicin,isepamicin, kanamycin, micronomicin, neomycin, neomycin undecylenate,netilmicin, paromomycin, ribostamycin, sisomicin, spectinomycin,streptomycin, tobramycin, trospectomycin), amphenicols (e.g.,azidamfenicol, chloramphenicol, florfenicol, thiamphenicol), ansamycins(e.g., rifamide, rifampin, rifamycin sv, rifapentine, rifaximin),P-lactams (e.g., carbacephems (e.g., loracarbef), carbapenems (e.g.,biapenem, imipenem, meropenem, panipenem), cephalosporins (e.g.,cefaclor, cefadroxil, cefamandole, cefatrizine, cefazedone, cefazolin,cefcapene pivoxil, cefclidin, cefdinir, cefditoren, cefepime, cefetamet,cefixime, cefinenoxime, cefodizime, cefonicid, cefoperazone, ceforanide,cefotaxime, cefotiam, cefozopran, cefpimizole, cefpiramide, cefpirome,cefpodoxime proxetil, cefprozil, cefroxadine, cefsulodin, ceftazidime,cefteram, ceftezole, ceftibuten, ceftizoxime, ceftriaxone, cefuroxime,cefuzonam, cephacetrile sodium, cephalexin, cephaloglycin,cephaloridine, cephalosporin, cephalothin, cephapirin sodium,cephradine, pivcefalexin), cephamycins (e.g., cefbuperazone,cefinetazole, cefminox, cefotetan, cefoxitin), monobactams (e.g.,aztreonam, carumonam, tigemonam), oxacephems, flomoxef, moxalactam),penicillins (e.g., amdinocillin, amdinocillin pivoxil, amoxicillin,ampicillin, apalcillin, aspoxicillin, azidocillin, azlocillin,bacampicillin, benzylpenicillinic acid, benzylpenicillin sodium,carbenicillin, carindacillin, clometocillin, cloxacillin, cyclacillin,dicloxacillin, epicillin, fenbenicillin, floxacillin, hetacillin,lenampicillin, metampicillin, methicillin sodium, mezlocillin, nafcillinsodium, oxacillin, penamecillin, penethamate hydriodide, penicillin gbenethamine, penicillin g benzathine, penicillin g benzhydrylamine,penicillin g calcium, penicillin g hydrabamine, penicillin g potassium,penicillin g procaine, penicillin n, penicillin o, penicillin v,penicillin v benzathine, penicillin v hydrabamine, penimepicycline,phenethicillin potassium, piperacillin, pivampicillin, propicillin,quinacillin, sulbenicillin, sultamicillin, talampicillin, temocillin,ticarcillin), ritipenem, lincosamides (e.g., clindamycin, lincomycin),macrolides (e.g., azithromycin, carbomycin, clarithromycin,dirithromycin, erythromycin, erythromycin acistrate, erythromycinestolate, erythromycin glucoheptonate, erythromycin lactobionate,erythromycin propionate, erythromycin stearate, josamycin, leucomycins,midecamycins, miokamycin, oleandomycin, primycin, rokitamycin,rosaramicin, roxithromycin, spiramycin, troleandomycin), polypeptides(e.g., amphomycin, bacitracin, capreomycin, colistin, enduracidin,enviomycin, fusafungine, gramicidin s, gramicidin(s), mikamycin,polymyxin, pristinamycin, ristocetin, teicoplanin, thiostrepton,tuberactinomycin, tyrocidine, tyrothricin, vancomycin, viomycin,virginiamycin, zinc bacitracin), tetracyclines (e.g., apicycline,chlortetracycline, clomocycline, demeclocycline, doxycycline,guamecycline, lymecycline, meclocycline, methacycline, minocycline,oxytetracycline, penimepicycline, pipacycline, rolitetracycline,sancycline, tetracycline), and others (e.g., cycloserine, mupirocin,tuberin); synthetic antibacterials, 2,4-Diaminopyrimidines (e.g.,brodimoprim, tetroxoprim, trimethoprim), nitrofurans (e.g., furaltadone,furazolium chloride, nifuradene, nifuratel, nifurfoline, nifurpirinol,nifurprazine, nifurtoinol, nitrofurantoin), quinolones and analogs(e.g., cinoxacin, ciprofloxacin, clinafloxacin, difloxacin, enoxacin,fleroxacin, flumequine, grepafloxacin, lomefloxacin, miloxacin,nadifloxacin, nalidixic acid, norfloxacin, ofloxacin, oxolinic acid,pazufloxacin, pefloxacin, pipemidic acid, piromidic acid, rosoxacin,rufloxacin, sparfloxacin, temafloxacin, tosufloxacin, trovafloxacin),sulfonamides (e.g., acetyl sulfamethoxypyrazine, benzylsulfamide,chloramine-b, chloramine-t, dichloramine t, n2-formylsulfisomidine,n4-β-d-glucosylsulfanilamide, mafenide,4′-(methylsulfamoyl)sulfanilanilide, noprylsulfamide,phthalylsulfacetamide, phthalylsulfathiazole, salazosulfadimidine,succinylsulfathiazole, sulfabenzamide, sulfacetamide,sulfachlorpyridazine, sulfachrysoidine, sulfacytine, sulfadiazine,sulfadicramide, sulfadimethoxine, sulfadoxine, sulfaethidole,sulfaguanidine, sulfaguanol, sulfalene, sulfaloxic acid, sulfamerazine,sulfameter, sulfamethazine, sulfamethizole, sulfamethomidine,sulfamethoxazole, sulfamethoxypyridazine, sulfametrole,sulfamidochrysoidine, sulfamoxole, sulfanilamide,4-sulfanilamidosalicylic acid, n4-sulfanilylsulfanilamide,sulfanilylurea, n-sulfanilyl-3,4-xylamide, sulfanitran, sulfaperine,sulfaphenazole, sulfaproxyline, sulfapyrazine, sulfapyridine,sulfasomizole, sulfasymazine, sulfathiazole, sulfathiourea,sulfatolamide, sulfisomidine, sulfisoxazole) sulfones (e.g., acedapsone,acediasulfone, acetosulfone sodium, dapsone, diathymosulfone,glucosulfone sodium, solasulfone, succisulfone, sulfanilic acid,p-sulfanilylbenzylamine, sulfoxone sodium, thiazolsulfone), and others(e.g., clofoctol, hexedine, methenamine, methenamineanhydromethylene-citrate, methenamine hippurate, methenamine mandelate,methenamine sulfosalicylate, nitroxoline, taurolidine, xibomol),antifungal antibiotics, polyenes (e.g., amphotericin b, candicidin,dermostatin, filipin, fungichromin, hachimycin, hamycin, lucensomycin,mepartricin, natamycin, nystatin, pecilocin, perimycin), azaserine,griseofulvin, oligomycins, neomycin undecylenate, pyrrolnitrin,siccanin, tubercidin, viridin, synthetic antifungals, allylamines (e.g.,butenafine, naftifine, terbinafine), imidazoles (e.g., bifonazole,butoconazole, chlordantoin, chlormidazole, cloconazole, clotrimazole,econazole, enilconazole, fenticonazole, flutrimazole, isoconazole,ketoconazole, lanoconazole, miconazole, omoconazole, oxiconazolenitrate, sertaconazole, sulconazole, tioconazole), thiocarbamates (e.g.,tolciclate, tolindate, tolnaftate), triazoles (e.g., fluconazole,itraconazole, saperconazole, terconazole), acrisorcin, amorolfine,biphenamine, bromosalicylchloranilide, buclosamide, calcium propionate,chlorphenesin, ciclopirox, cloxyquin, coparaffinate, diamthazoledihydrochloride, exalamide, flucytosine, halethazole, hexetidine,loflucarban, nifuratel, potassium iodide, propionic acid, pyrithione,salicylanilide, sodium propionate, sulbentine, tenonitrozole, triacetin,ujothion, undecylenic acid, zinc propionate, antineoplastics,antibiotics and analogs (e.g., aclacinomycins, actinomycin f1,anthramycin, azaserine, bleomycins, cactinomycin, carubicin,carzinophilin, chromomycins, dactinomycin, daunorubicin,6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, idarubicin,menogaril, mitomycins, mycophenolic acid, nogalamycin, olivomycines,peplomycin, pirarubicin, plicamycin, porfiromycin, puromycin,streptonigrin, streptozocin, tubercidin, zinostatin, zorubicin),antimetabolites (e.g. folic acid analogs (e.g., denopterin, edatrexate,methotrexate, piritrexim, pteropterin, Tomudex®, trimetrexate), purineanalogs (e.g., cladribine, fludarabine, 6-mercaptopurine, thiamiprine,thioguanine), pyrimidine analogs (e.g., ancitabine, azacitidine,6-azauridine, carmofur, cytarabine, doxifluridine, emitefur,enocitabine, floxuridine, fluorouracil, gemcitabine, tagafur),antiinflammatory agents, steroidal antiinflammatory agents,acetoxypregnenolone, alclometasone, algestone, amcinonide,beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol,clobetasone, clocortolone, cloprednol, corticosterone, cortisone,cortivazol, deflazacort, desonide, desoximetasone, dexamethasone,diflorasone, diflucortolone, difluprednate, enoxolone, fluazacort,flucloronide, flumethasone, flunisolide, fluocinolone acetonide,fluocinonide, fluocortin butyl, fluocortolone, fluorometholone,fluperolone acetate, fluprednidene acetate, fluprednisolone,flurandrenolide, fluticasone propionate, formocortal, halcinonide,halobetasol propionate, halometasone, halopredone acetate,hydrocortamate, hydrocortisone, loteprednol etabonate, mazipredone,medrysone, meprednisone, methylprednisolone, mometasone furoate,paramethasone, prednicarbate, prednisolone, prednisolone25-diethylamino-acetate, prednisolone sodium phosphate, prednisone,prednival, prednylidene, rimexolone, tixocortol, triamcinolone,triamcinolone acetonide, triamcinolone benetonide, and triamcinolonehexacetonide, non-steroidal antiinflammatory agents, aminoarylcarboxylicacid derivatives (e.g., enfenamic acid, etofenamate, flufenamic acid,isonixin, meclofenamic acid, mefenamic acid, niflumic acid,talniflumate, terofenamate, tolfenamic acid), arylacetic acidderivatives (e.g., aceclofenac, acemetacin, alclofenac, amfenac,amtolmetin guacil, bromfenac, bufexamac, cinmetacin, clopirac,diclofenac sodium, etodolac, felbinac, fenclozic acid, fentiazac,glucametacin, ibufenac, indomethacin, isofezolac, isoxepac, lonazolac,metiazinic acid, mofezolac, oxametacine, pirazolac, proglumetacin,sulindac, tiaramide, tolmetin, tropesin, zomepirac), arylbutyric acidderivatives (e.g., bumadizon, butibufen, fenbufen, xenbucin),arylcarboxylic acids (e.g., clidanac, ketorolac, tinoridine),arylpropionic acid derivatives (e.g., alminoprofen, benoxaprofen,bermoprofen, bucloxic acid, carprofen, fenoprofen, flunoxaprofen,flurbiprofen, ibuprofen, ibuproxam, indoprofen, ketoprofen, loxoprofen,naproxen, oxaprozin, piketoprolen, pirprofen, pranoprofen, protizinicacid, suprofen, tiaprofenic acid, ximoprofen, zaltoprofen), pyrazoles(e.g., difenamizole, epirizole), pyrazolones (e.g., apazone,benzpiperylon, feprazone, mofebutazone, morazone, oxyphenbutazone,phenylbutazone, pipebuzone, propyphenazone, ramifenazone, suxibuzone,thiazolinobutazone), salicylic acid derivatives (e.g., acetaminosalol,aspirin, benorylate, bromosaligenin, calcium acetylsalicylate,diflunisal, etersalate, fendosal, gentisic acid, glycol salicylate,imidazole salicylate, lysine acetylsalicylate, mesalamine, morpholinesalicylate, 1-naphthyl salicylate, olsalazine, parsalmide, phenylacetylsalicylate, phenyl salicylate, salacetamide, salicylamide o-aceticacid, salicylsulfuric acid, salsalate, sulfasalazine),thiazinecarboxamides (e.g., ampiroxicam, droxicam, isoxicam, lomoxicam,piroxicam, tenoxicam), β-acetamidocaproic acid, s-adenosylmethionine,3-amino-4-hydroxybutyric acid, amixetrine, bendazac, benzydamine,a-bisabolol, bucolome, difenpiramide, ditazol, emorfazone, fepradinol,guaiazulene, nabumetone, nimesulide, oxaceprol, paranyline, perisoxal,proquazone, superoxide dismutase, tenidap, and zileuton.

The therapeutic agents may also be used in combination with othertherapeutic agents and therapies, including but not limited to agentsand therapies useful for the treatment, prevention, inhibition, delayingonset of, or causing regression of angiogenesis or neovascularization,particularly CNV. In some variations the additional agent or therapy isused to treat regression of angiogenesis or neovascularization,particularly CNV. Non-limiting examples of such additional agents andtherapies include pyrrolidine, dithiocarbamate (NFκB inhibitor);squalamine; TPN 470 analogue and fumagillin; PKC (protein kinase C)inhibitors; Tie-1 and Tie-2 kinase inhibitors; inhibitors of VEGFreceptor kinase; proteosome inhibitors such as Velcade™ (bortezomib, forinjection; ranibuzumab (Lucentis™) and other antibodies directed to thesame target; pegaptanib (Macugen™); vitronectin receptor antagonists,such as cyclic peptide antagonists of vitronectin receptor-typeintegrins; α-v/β-3 integrin antagonists; α-v/β-1 integrin antagonists;thiazolidinediones such as rosiglitazone or troglitazone; interferon,including γ-interferon or interferon targeted to CNV by use of dextranand metal coordination; pigment epithelium derived factor (PEDF);endostatin; angiostatin; tumistatin; canstatin; anecortave acetate;acetonide; triamcinolone; tetrathiomolybdate; RNA silencing or RNAinterference (RNAi) of angiogenic factors, including ribozymes thattarget VEGF expression; Accutane™ (13-cis retinoic acid); ACEinhibitors, including but not limited to quinopril, captopril, andperindozril; inhibitors of mTOR (mammalian target of rapamycin);3-aminothalidomide; pentoxifylline; 2-methoxyestradiol; colchicines;AMG-1470; cyclooxygenase inhibitors such as nepafenac, rofecoxib,diclofenac, rofecoxib, NS398, celecoxib, vioxx, and(E)-2-alkyl-2(4-methanesulfonylphenyl)-1-phenylethene; t-RNA synthasemodulator; metalloprotease 13 inhibitor; acetylcholinesterase inhibitor;potassium channel blockers; endorepellin; purine analog of6-thioguanine; cyclic peroxide ANO-2; (recombinant) arginine deiminase;epigallocatechin-3-gallate; cerivastatin; analogues of suramin; VEGFtrap molecules; inhibitors of hepatocyte growth factor (antibodies tothe growth factor or its receptors, small molecular inhibitors of thec-met tyrosine kinase, truncated versions of HGF e.g. NK4); apoptosisinhibiting agents; Visudyne™, snET2 and other photo sensitizers withphotodynamic therapy (PDT); and laser photocoagulation.

In some variations, the therapeutic agent is used in combination withother therapeutic agents. In some variations the therapeutic agent isused in combination with verteporfin (Visudyne™). In some variations thetherapeutic agent is used in combination with an inhibitor of VascularEndothelial Growth Factor (VEGF). In some variations the therapeuticagent is used in combination with an inhibitor of Vascular EndothelialGrowth Factor-A (VEGF-A). In some variations, the inhibitor of VEGF is aVEGF trap molecule including, without limitation, the VEGF trap underdevelopment by Regeneron Pharmaceuticals. In some variations theinhibitor of VEGF is an antibody or fragment thereof directed to VEGF.In some variations the antibody or fragment thereof directed to VEGF isbevacizumab (i.e., Avastin™). In some variations the antibody orfragment thereof directed to VEGF is ranibizumab (i.e., Lucentis™). Insome variations, the therapeutic agent may be a limus compound. In somevariations, the therapeutic agent may be rapamycin or an analog orderivative thereof including those described in the Therapeutic Agentssection. In some variations, the therapeutic agent is rapamycin. In somevariations, the therapeutic agent is dasatinib.

In some variations, the therapeutic agent is a limus compound such asrapamycin or an analog or derivative thereof including those describedin the Therapeutic Agents section and an other therapeutic agent such asranibizumab (i.e., Lucentis™), bevacizumab (i.e., Avastin™), or a VEGFtrap. In some variations, the therapeutic agent is a limus compound suchas rapamycin or an analog or derivative thereof including thosedescribed in the Therapeutic Agents section and the other therapeuticagent is ranibizumab (i.e., Lucentis™) or bevacizumab (i.e., Avastin™).Also provided are methods for treating or preventing wet-AMD in a humansubject comprising administration of a liquid formulation comprising atherapeutic agent such as rapamycin or an analog or derivative thereofincluding those described in the Therapeutic Agents section and an othertherapeutic agent such as ranibizumab (i.e., Lucentis™), bevacizumab(i.e., Avastin™), or a VEGF trap. In some variations, the methods fortreating or preventing wet-AMD in a human subject comprisingadministration of a liquid formulation comprising rapamycin andranibizumab (i.e., Lucentis™), bevacizumab (i.e., Avastin™), or a VEGFtrap. In some variations, the liquid formulation comprising rapamycinand ranibizumab (i.e., Lucentis™), bevacizumab (i.e., Avastin™), or aVEGF trap is administered intravitreally, subconjunctivally,subtenonally, or topically. In some variations, the liquid formulationis administered intravitreally. In some variations, the total amount ofLucentis or Avastin is any of about 0.1 mg, about 0.2 mg, about 0.3 mg,about 0.4 mg, about 0.5 mg, about 0.6 mg, or about 0.7 mg. In somevariations, the total amount of Lucentis or Avastin is 0.3 mg or 0.5 mg.

In some variations the therapeutic agent used in combination with theother therapeutic agent may be administered simultaneously (i.e.,simultaneous administration), sequentially (i.e., sequentialadministration), or independently of the other therapeutic agents. Insome variations the therapeutic agent and the other therapeutic agentare administered by the same route of administration. In somevariations, the therapeutic agent and other therapeutic agent are bothadministered during one visit to the physician. In some variations, thetherapeutic agent and other therapeutic agent are both administeredduring the same physician visit. In some variations, the therapeuticagent and other therapeutic agent are administered during more than onevisit to the physician.

In some variations the therapeutic agent and the other therapeutic agentare administered by the same route of administration. In some variationsthe therapeutic agent and the other therapeutic agent are administeredby different routes of administration. In some variations thetherapeutic agent are administered by subconjunctival administrationincluding, without limitation, subconjunctival injection, and the othertherapeutic agent are administered by intravitreal administrationincluding, without limitation, intravitreal injection. In somevariations the therapeutic agent are administered by intravitrealadministration including, without limitation, intravitreal injection,and the other therapeutic agent are administered by subconjunctivaladministration including, without limitation, subconjunctival injection.In some variations the therapeutic agent are administered bysubconjunctival administration including, without limitation,subconjunctival injection, and the other therapeutic agent areadministered by subtenon administration including, without limitation,subtenon injection. In some variations the therapeutic agent areadministered by subtenon administration including, without limitation,subtenon injection, and the other therapeutic agent are administered bysubconjunctival administration including, without limitation,subconjunctival injection. In some variations the therapeutic agent areadministered by intravitreal administration including, withoutlimitation, intravitreal injection, and the other therapeutic agent areadministered by subtenon administration including, without limitation,subtenon injection. In some variations the therapeutic agent areadministered by subtenon administration including, without limitation,subtenon injection, and the other therapeutic agent are administered byintravitreal administration including, without limitation, intravitrealinjection. In some variations, the therapeutic agent is administeredtopically and the other therapeutic agent is administered bysubconjunctival, subtenon, and/or intravitreal administration. In somevariations, the other therapeutic agent is administered topically andthe therapeutic agent is administered by subconjunctival, subtenon,and/or intravitreal administration. In some variations the formulationof the therapeutic agent and the other therapeutic agent are the same(i.e., identical).

In some variations the combination of administration of therapeuticagent and administration of other therapeutic agent reduces the doseadministered (e.g., dose volume, dose concentration, or amount oftherapeutic agent) of one or both therapeutic agents. In some variationsthe combination of therapeutic agent and other therapeutic agentincreases or prolongs the time between administrations and/or decreasesthe frequency of administrations of one or both therapeutic agents.

In some variations, the therapeutic agent is rapamycin. In somevariations, the therapeutic agent is dasatinib. In some variations, theother therapeutic agent is verteporfin, bevacizumab, and/or ranibizumab.

By way of non-limiting example, administration of rapamycin may be usedto decrease the frequency of administration of bevacizumab orranibizumab. In one non-limiting example, wet AMD is treated byadministering one, two, or three doses of Avastin or Lucentis followedby subsequent doses of rapamycin, alone or in combination with Avastinor Lucentis.

Diseases and Conditions that May be Treated, Prevented, Inhibited, OnsetDelayed, or Regression Caused

Herein are described diseases and conditions that may be treated,prevented, inhibited, onset delayed, or regression caused using thetherapeutic agents and the formulations, liquid formulations, andmethods described herein. In some variations, the diseases or conditionsare treated using the therapeutic agents and the formulations, liquidformulations, and methods described herein. Unless the context indicatesotherwise, it is envisioned that the subjects on whom all of the methodsof treatment may be performed include, but are not limited to, humansubjects.

Generally, any diseases or condition of the eye susceptible totreatment, prevention, inhibition, delaying the onset of, or causing theregression of using the therapeutic agents and the formulations, liquidformulations and methods described herein may be treated, prevented,inhibited, onset delayed, or regression caused treated or prevented.Examples of diseases or conditions of the eye include, but are notlimited to, diseases or conditions associated with neovascularizationincluding retinal and/or choroidal neovascularization.

Diseases or conditions associated with retinal and/or choroidalneovascularization that can be treated, prevented inhibited, have onsetdelayed, or be caused to regress using the formulations, liquidformulations, and methods described herein include, but are not limitedto, diabetic retinopathy, macular degeneration, wet and dry AMD,retinopathy of prematurity (retrolental fibroplasia), infections causinga retinitis or choroiditis, presumed ocular histoplasmosis, myopicdegeneration, angioid streaks, and ocular trauma. Other non-limitingexamples of diseases and conditions of the eye that may be treated,prevented inhibited, have onset delayed, or be caused to regress usingthe formulations, liquid formulations, and methods described hereininclude, but are not limited to, pseudoxanthoma elasticum, veinocclusion, artery occlusion, carotid obstructive disease, Sickle Cellanemia, Eales disease, myopia, chronic retinal detachment,hyperviscosity syndromes, toxoplasmosis, trauma, polypoidal choroidalvasculopathy, post-laser complications, complications of idiopathiccentral serous chorioretinopathy, complications of choroidalinflammatory conditions, rubeosis, diseases associated with rubeosis(neovascularization of the angle), neovascular glaucoma, uveitis andchronic uveitis, macular edema, proliferative retinopathies and diseasesor conditions caused by the abnormal proliferation of fibrovascular orfibrous tissue, including all forms of proliferative vitreoretinopathy(including post-operative proliferative vitreoretinopathy), whether ornot associated with diabetes.

In some variations, the formulations and pharmaceutical formulationsdescribed herein are used to prevent or delay onset of a disease orcondition of the eye where the subject, including but not limited to ahuman subject, is at heightened risk of developing the disease orcondition of the eye. A subject with a heightened risk of developing adisease or condition is a subject with one or more indications that thedisease or condition is likely to develop in the particular subject. Insome variations the subject with a heightened risk of developing wet AMDis a subject with dry AMD in at least one eye. In some variations thesubject with a heightened risk of developing wet AMD in a fellow eye isa subject with wet AMD in the other eye. In some variations, theformulations and pharmaceutical formulations described herein are usedto prevent or delay onset of CNV in a subject at heightened risk ofdeveloping CNV, including but not limited to prevention or delayingonset of CNV in the fellow eye of a subject, including but not limitedto a human subject with AMD in one eye. In some variations, theformulations and pharmaceutical formulations described herein are usedto prevent or delay onset of CNV in the fellow eye of a subject with wetAMD in one eye. In some variations, the formulations and pharmaceuticalformulations comprise a limus compound, including but not limited torapamycin. In some variations the formulations and pharmaceuticalformulations are administered periocularly, including without limitationsubconjunctivally, to a human subject with vision of 20/40 or better. Insome variations, the formulations and pharmaceutical formulations areadministered periocularly, including without limitationsubconjunctivally, to the eye of a human subject where the eye to whichthe formulation is administered has vision of 20/40 or better.

In some variations, the formulations and pharmaceutical formulationsdescribed herein are used to treat, prevent, or delay onset of AMD. Insome variations, the formulations and pharmaceutical formulationsdescribed herein are used to treat, prevent, or delay onset of dry AMD.In some variations, subjects including but not limited to human subjectswith non-central geographic atrophy are administered a formulation orpharmaceutical formulations described herein to treat, prevent, or delayonset of central geographic atrophy. In some variations, theformulations and pharmaceutical formulations comprise a limus compound,including but not limited to rapamycin. In some variations theformulations and pharmaceutical formulations are administeredperiocularly, including without limitation subconjunctivally, to a humansubject with vision of 20/40 or better. In some variations, theformulations and pharmaceutical formulations described herein areadministered and the subject, including but not limited to a humansubject is also treated with a second therapy for treating the diseaseor disorder. In some variations, the formulations and pharmaceuticalformulations described herein are used to treat, prevent, or delay onsetof wet or dry AMD and the subject, including but not limited to a humansubject is also treated with laser therapy such as photodynamic lasertherapy, either before, during, or after treatment with the formulationsor pharmaceutical formulations described herein.

In some variations, the formulations and pharmaceutical formulationsdescribed herein are used to treat one or more of uveitis, allergicconjunctivitis, macular edema, glaucoma, or dry eye.

In some variations, a formulations or pharmaceutical formulationcomprises a limus compound such as rapamycin, and is administered totreat, prevent, or delay onset of dry eye. In some variations, aformulations or pharmaceutical formulation comprises a limus compoundsuch as rapamycin, and is administered to treat, prevent, or delay onsetof allergic conjunctivitis.

In some variations, the formulations and pharmaceutical formulationsdescribed herein are used to treat glaucoma. In some variations, theformulations and pharmaceutical formulations described herein fortreating glaucoma comprise a limus compound such as rapamycin, and areused as a surgical adjuvant to prevent, reduce or delay surgicalcomplications. In some variations, the formulations and pharmaceuticalformulations described herein for treating glaucoma comprise a limuscompound such as rapamycin, and are used to improve or prolong surgicalimplant success. In some variations, the formulations and pharmaceuticalformulations described herein for treating glaucoma comprise a limuscompound such as rapamycin, and are used to improve or prolong successof an argon laser trabeculectomy or other glaucoma-related surgery. Insome variations, the formulations and pharmaceutical formulationsdescribed herein have a neuroprotective effect and are used to treatglaucoma.

In some variations, the formulations and pharmaceutical formulationsdescribed herein are used to treat retinitis pigmentosa. In somevariations, the formulations and pharmaceutical formulations describedherein for treating glaucoma comprise a limus compound such asrapamycin, and are used to treat, prevent, or delay onset of retinitispigmentosa. In some variations, the formulations and pharmaceuticalformulations described herein have a neuroprotective effect and are usedto treat retinitis pigmentosa.

In some variations, the formulations and pharmaceutical formulationsdescribed herein are used to treat one or more of central retinal veinocclusive diseases (CRVO), branch retinal venous occlusion (BRVO),retinal vascular diseases and conditions, macular edema, diabeticmacular edema, iris neovascularization, diabetic retinopathy, or cornealgraft rejection. In some variations, a formulations or pharmaceuticalformulation comprises a limus compound such as rapamycin, and isadministered to treat, prevent, or delay onset of one or more of thesediseases or conditions. In some variations the formulations andpharmaceutical formulations are administered subconjunctivally to an eyewith vision of 20/40 or better.

When used to treat, prevent, inhibit, delay the onset of, or causeregressions of uveitis, the formulations and pharmaceutical formulationsdescribed herein may be administered by a variety of routes as is knownin the art, including but not limited to by ocular or oraladministration. Other routes of administration are known and are routinein the art. In some variations, the formulations described hereincomprise rapamycin and are used to treat uveitis.

One disease that may be treated, prevented, inhibited, have onsetdelayed, or be caused to regress using the formulation, liquidformulations and methods described herein is the wet form of AMD. Insome variations wet AMD is treated using the formulations, liquidformulations and methods described herein. The wet form of AMD ischaracterized by blood vessels growing from their normal location in thechoroid into an undesirable position under the retina. Leakage andbleeding from these new blood vessels results in vision loss andpossibly blindness.

The formulations, liquid formulations, and methods described herein mayalso be used to prevent or slow the transition from the dry form of AMD(wherein the retinal pigment epithelium or RPE degenerates and leads tophotoreceptor cell death and the formation of yellow deposits calleddrusen under the retina) to the wet form of AMD.

“Macular degeneration” is characterized by the excessive buildup offibrous deposits in the macula and retina and the atrophy of the retinalpigment epithelium. As used herein, an eye “afflicted” with maculardegeneration is understood to mean that the eye exhibits at least onedetectable physical characteristic associated with the disease ofmacular degeneration. The administration of rapamycin appears to limitand regress angiogenesis, such as choroidal neovascularization inage-related macular degeneration (AMD), which may occur withouttreatment. As used herein, the term “angiogenesis” means the generationof new blood vessels (“neovascularization”) into a tissue or organ. An“angiogenesis-mediated disease or condition” of the eye or retina is onein which new blood vessels are generated in a pathogenic manner in theeye or retina, resulting in dimunition or loss of vision or otherproblem, e.g., choroidal neovascularization associated with AMD.

The formulations and liquid formulations described herein, including butnot limited to rapamycin-containing formulations and liquidformulations, may also be used to treat, prevent, inhibit, delay theonset of, or cause regression of various immune-related diseases andconditions, including but not limited to organ transplant rejection in ahost, graft vs. host disease, autoimmune diseases, diseases ofinflammation, hyperproliferative vascular disorders, solid tumors, andfungal infections. In some variations, the formulations and liquidformulations described herein, including but not limited torapamycin-containing formulations and liquid formulations, are used totreat various immune-related diseases and conditions, including but notlimited to organ transplant rejection in a host, graft vs. host disease,autoimmune diseases, diseases of inflammation, hyperproliferativevascular disorders, solid tumors, and fungal infections. Theformulations and liquid formulations described herein, including but notlimited to rapamycin-containing formulations and liquid formulations,may be used as immunosuppressants. The formulations and liquidformulations described herein, including but not limited torapamycin-containing formulations and liquid formulations, may be usedto treat, prevent, inhibit, or delay the onset of rejection oftransplanted organs or tissues including but not limited to transplantedheart, liver, kidney, spleen, lung, small bowel, pancreas, and bonemarrow. In some variations, the formulations and liquid formulationsdescribed herein are used to treat the onset of rejection oftransplanted organs or tissues including but not limited to transplantedheart, liver, kidney, spleen, lung, small bowel, pancreas, and bonemarrow. When used to treat, prevent, inhibit, delay the onset of, orcause regressions of immune-related diseases, including but not limitedto transplant rejection, the formulations and liquid formulationsdescribed herein may be administered by a variety of routes as is knownin the art, including but not limited to by oral administration.

Systemic administration may be achieved by oral administration of theliquid formulation. Other systemic routes of administration are knownand are routine in the art. Some examples thereof are listed in theDetailed Description section.

As used herein, to “inhibit” a disease or condition by administration ofa therapeutic agent means that the progress of at least one detectablephysical characteristic or symptom of the disease or condition is slowedor stopped following administration of the therapeutic agent as comparedto the progress of the disease or condition without administration ofthe therapeutic agent.

As used herein, to “prevent” a disease or condition by administration ofa therapeutic agent means that the detectable physical characteristicsor symptom of the disease or condition do not develop followingadministration of the therapeutic agent.

As used herein, to “delay onset of” a disease or condition byadministration of a therapeutic agent means that at least one detectablephysical characteristic or symptom of the disease or condition developslater in time following administration of the therapeutic agent ascompared to the progress of the disease or condition withoutadministration of the therapeutic agent.

As used herein, to “treat” a disease or condition by administration of atherapeutic agent means that the progress of at least one detectablephysical characteristic or symptom of the disease or condition isslowed, stopped, or reversed following administration of the therapeuticagent as compared to the progress of the disease or condition withoutadministration of the therapeutic agent.

As used herein, to “cause regression of” a disease or condition byadministration of a therapeutic agent means that the progress of atleast one detectable physical characteristic or symptom of the diseaseor condition is reversed to some extent following administration of thetherapeutic agent.

A subject, including but not limited to a human subject, having apredisposition for or in need of prevention may be identified by theskilled practitioner by established methods and criteria in the fieldgiven the teachings herein. The skilled practitioner may also readilydiagnose individuals as in need of inhibition or treatment based uponestablished criteria in the field for identifying angiogenesis and/orneovascularization given the teachings herein.

As used herein, a “subject” is generally any animal that may benefitfrom administration of the therapeutic agents described herein. In somevariations the therapeutic agents are administered to a mammaliansubject. In some variations the therapeutic agents are administered to ahuman subject. In some variations the therapeutic agents may beadministered to a veterinary animal subject. In some variations thetherapeutic agents may be administered to a model experimental animalsubject.

Other diseases and conditions that may be treated, prevented, inhibited,have the onset delayed, or be caused to regress using the methodsdescribed herein include those disclosed in the following patents andpublications, the contents of each of which is incorporated herein inits entirety: PCT publication WO 2004/027027, published Apr. 1, 2004,titled Method of inhibiting choroidal neovascularization, assigned toTrustees of the University of Pennsylvania; U.S. Pat. No. 5,387,589,issued Feb. 7, 1995, titled Method of Treating Ocular Inflammation, withinventor Prassad Kulkami, assigned to University of Louisville ResearchFoundation; U.S. Pat. No. 6,376,517, issued Apr. 23, 2003, titledPipecolic acid derivatives for vision and memory disorders, assigned toGPI NIL Holdings, Inc; PCT publication WO 2004/028477, published Apr. 8,2004, titled Method subretinal administration of therapeutics includingsteroids: method for localizing pharmadynamic action at the choroid andretina; and related methods for treatment and or prevention of retinaldiseases, assigned to Innorx, Inc; U.S. Pat. No. 6,416,777, issued Jul.9, 2002, titled Ophthalmic drug delivery device, assigned to AlconUniversal Ltd; U.S. Pat. No. 6,713,081, issued Mar. 30, 2004, titledOcular therapeutic agent delivery device and methods for making andusing such devices, assigned to Department of Health and Human Services;and U.S. Pat. No. 5,536,729, issued Jul. 16, 1996, titled RapamycinFormulations for Oral Administration, assigned to American Home ProductsCorp., and U.S. Pat. App. Nos. 60/503,840 and 10/945,682.

Liquid Formulations

The liquid formulations described herein contain a therapeutic agent andmay generally be any liquid formulation, including but not limited tosolutions, suspensions, and emulsions. In some variations the liquidformulations form a non-dispersed mass relative to a surrounding mediumwhen placed in the vitreous of a rabbit eye.

When a certain volume of a liquid formulation is administered, it isunderstood that there is some imprecision in the accuracy of variousdevices that may be used to administer the liquid formulation. Where acertain volume is specified, it is understood that this is the targetvolume. However, certain devices such as insulin syringes are inaccurateto greater than 10%, and sometimes inaccurate to greater than 20% ormore. Hamilton HPLC type syringes are generally considered precise towithin 10%, and are recommended for volumes below 10 μl are to beinjected.

In some variations, a volume of a liquid formulation described herein isadministered to the vitreous of a rabbit eye or a subject's, includingbut not limiting a human subject's eye that is less than about 500 μl,less than about 400 μl, less than about 300 μl, less than about 200 μl,less than about 100 μl, less than about 90 μl, less than about 80 μl,less than about 70 μl, less than about 60 μl, less than about 50 μl,less than about 40 μl, less than about 30 μl, less than about 20 μl,less than about 10 μl, less than about 5 μl, less than about 3 μl, orless than about 1 μl. In some variations, a volume of a liquidformulation described herein is administered to the vitreous of a rabbiteye or subject's, including but not limited to a human subject's eyethat is less than about 20 μl. In some variations, a volume of a liquidformulation described herein is administered to the vitreous that isless than about 10 μl. In some variations, a volume of a liquidformulation described herein is administered to the vitreous of a rabbiteye or a subject's, including but not limited to a human subject's eyethat is between about 0.1 μl and about 200 μl, between about 50 μl andabout 200 μl, between about 50 μl and about 150 μl, between about 0.1 μland about 100 μl, between about 0.1 μl and about 50 μl, between about 1μl and about 40 μl, between about 1 μl and about 30 μl, between about 1μl and about 20 μl, between about 1 μl and about 10 μl, or between about1 μl and about 5 μl. In some variations, a volume of a liquidformulation described herein is administered to the vitreous of a rabbiteye or a subject's, including but not limited to a human subject's eyethat is between about 1 μl and about 10 μl. In some variations, a volumeof a liquid formulation described herein is administered to the vitreousof a rabbit eye or a subject's, including but not limited to a humansubject's eye that is between about 1 μl and about 5 μl. In somevariations, a volume of a liquid formulation described herein isadministered to the vitreous of a rabbit eye or a subject's eye that isbetween about 11 and about 5 μl. In some variations, a volume of aliquid formulation described herein is administered to the vitreous of arabbit eye or a subject's, including but not limited to a humansubject's eye that is between about 0.1 μl and about 200 μl.

In some variations, a total volume of a liquid formulation describedherein is subconjunctivally administered to a rabbit eye or a subject's,including but not limited to a human subject's eye that is less thanabout 1000 μl, less than about 900 μl, less than about 800 μl, less thanabout 700 μl, less than about 600 μl, less than about 500 μl, less thanabout 400 μl, less than about 300 μl, less than about 200 μl, less thanabout 100 μl, less than about 90 μl, less than about 80 μl, less thanabout 70 μl, less than about 60 μl, less than about 50 μl, less thanabout 40 μl, less than about 30 μl, less than about 20 μl, less thanabout 10 μl, less than about 5 μl, less than about 3 μl, or less thanabout 1 μl. In some variations, a volume of a liquid formulationdescribed herein is subconjunctivally administered to a rabbit eye or asubject's, including but not limited to a human subject's eye that isless than about 20 μl. In some variations, a volume of a liquidformulation described herein is subconjunctivally administered to arabbit eye or a subject's, including but not limited to a humansubject's eye that is less than about 10 μl. In some variations, avolume of a liquid formulation described herein is subconjunctivallyadministered to a rabbit eye or a subject's, including but not limitedto a human subject's eye that is between about 0.11 and about 200 μl,between about 50 μl and about 200 μl, between about 200 μl and about 300μl, between about 300 μl and about 400 μl, between about 400 μl andabout 500 μl, between about 600 μl and about 700 μl, between about 700μl and about 800 μl, between about 800 μl and about 900 μl, betweenabout 900 μl and about 1000 μl, between about 50 μl and about 150 μl,between about 0.1 μl and about 100 μl, between about 0.1 μl and about 50μl, between about 1 μl and about 40 μl, between about 1 μl and about 30μl, between about 1 μl and about 20 μl, between about 1 μl and about 10μl, or between about 1 μl and about 5 μl. In some variations, a volumeof a liquid formulation described herein is subconjunctivallyadministered to a rabbit eye or a subject's, including but not limitedto a human subject's eye that is between about 1 μl and about 10 μl. Insome variations, a volume of a liquid formulation described herein issubconjunctivally administered to a rabbit eye or a subject's, includingbut not limited to a human subject's eye that is between about 1 μl andabout 5 μl. In some variations, a volume of a liquid formulationdescribed herein is administered to subconjunctivally administered to arabbit eye or a subject's, including but not limited to a humansubject's eye that is between about 1 μl and about 5 μl. In somevariations, a volume of a liquid formulation described herein isadministered to subconjunctivally administered to a rabbit eye or asubject's, including but not limited to a human subject's eye that isbetween about 0.1 μl and about 200 μl.

In some variations, a volume of a liquid formulation described herein isadministered to the vitreous of a rabbit eye or a subject's, includingbut not limited to a human subject's eye that is any of about 2 μl,about 4 μl, about 5 μl, about 6 μl, about 8 μl, about 10 μl, about 12μl, about 14 μl, about 16 μl, about 18 μl, or about 20 μl. In somevariations, a volume of a liquid formulation described herein isadministered to the vitreous of a rabbit eye or a subject's, includingbut not limited to a human subject's eye that is about 2 μl. In somevariations, a volume of a liquid formulation described herein isadministered to the vitreous of a rabbit eye or a subject's, includingbut not limited to a human subject's eye that is about 5 μl. In somevariations, a volume of a liquid formulation described herein isadministered to the vitreous of a rabbit eye or a subject's, includingbut not limited to a human subject's eye that is about 8 μl.

In some variations, a total volume of a liquid formulation issubconjunctivally administered to a rabbit eye or a subject's, includingbut not limited to a human subject's eye that is less than about 150 μl.In some variations, a volume of a liquid formulation described herein issubconjunctivally administered to a rabbit eye or a subject's, includingbut not limited to a human subject's eye that is any of about 10 μl,about 20 μl, about 30 μl, about 40 μl, about 50 μl, about 60 μl, about70 μl, about 80 μl, about 90 μl, or about 100 μl. In some variations, avolume of a liquid formulation described herein is subconjunctivallyadministered to a rabbit eye or a subject's, including but not limitedto a human subject's eye that is about 10 μl. In some variations, avolume of a liquid formulation described herein is subconjunctivallyadministered to a rabbit eye or a subject's, including but not limitedto a human subject's eye that is about 20 μl. In some variations, avolume of a liquid formulation described herein is subconjunctivallyadministered to a rabbit eye or a subject's, including but not limitedto a human subject's eye that is about 30 μl. In some variations, avolume of a liquid formulation described herein is subconjunctivallyadministered to a rabbit eye or a subject's, including but not limitedto a human subject's eye that is 40 μl. In some variations, a volume ofa liquid formulation described herein is subconjunctivally administeredto a rabbit eye or a subject's, including but not limited to a humansubject's eye that is any of between about 10 μl and about 50 μl,between about 15 μl and about 45 μl, between about 20 μl and about 40μl, or between about 25 μl and about 35 μl.

In some variations, a total volume of a liquid formulation describedherein is subtenonally administered to a rabbit eye or a subject's,including but not limited to a human subject's eye that is less thanabout 1000 μl, less than about 900 μl, less than about 800 μl, less thanabout 700 μl, less than about 600 μl, less than about 500 μl, less thanabout 400 μl, less than about 300 μl, less than about 200 μl, less thanabout 150 μl, less than about 100 μl, less than about 90 μl, less thanabout 80 μl, less than about 70 μl, less than about 60 μl, less thanabout 50 μl, less than about 40 μl, less than about 30 μl, less thanabout 20 μl, less than about 10 μl, less than about 5 μl, less thanabout 3 μl, or less than about 1 μl. In some variations, a volume of aliquid formulation described herein is subtenonally administered to arabbit eye or a subject's, including but not limited to a humansubject's eye that is less than about 200 μl. In some variations, avolume of a liquid formulation described herein is subtenonallyadministered to a rabbit eye or a subject's, including but not limitedto a human subject's eye that is less than about 100 μl. In somevariations, a volume of a liquid formulation described herein issubtenonally administered to a rabbit eye or a subject's, including butnot limited to a human subject's eye that is between about 0.1 μl andabout 200 pt, between about 50 μl and about 200 μl, between about 200 μland about 300 μl, between about 300 μl and about 400 μl, between about400 μl and about 500 μl, between about 600 μl and about 700 μl, betweenabout 700 μl and about 800 μl, between about 800 μl and about 900 μl,between about 900 μl and about 1000 μl, between about 50 μl and about150 μl, between about 0.1 μl and about 100 μl, between about 0.1 μl andabout 50 μl, between about 1 μl and about 40 μl, between about 1 μl andabout 30 μl, between about 1 μl and about 20 μl, between about 1 μl andabout 10 μl, or between about 1 μl and about 5 μl. In some variations, avolume of a liquid formulation described herein is subtenonallyadministered to a rabbit eye or a subject's, including but not limitedto a human subject's eye that is between about 10 μl and about 200 μl.In some variations, a volume of a liquid formulation described herein isadministered subtenonally to a rabbit eye or a subject's, including butnot limited to a human subject's eye that is between about 0.1 μl andabout 200 μl. In some variations, a volume of a liquid formulationdescribed herein is subtenonally administered to a rabbit eye or asubject's, including but not limited to a human subject's eye that isbetween about 50 μl and about 150 μl. In some variations, a volume of aliquid formulation described herein is subtenonally administered to arabbit eye or a subject's, including but not limited to a humansubject's eye that is about 30 μl. In some variations, a volume of aliquid formulation described herein is subtenonally administered to arabbit eye or a subject's, including but not limited to a humansubject's eye that is about 120 μl.

In some variations, a total amount of therapeutic agent less than about5 mg is administered subtenonally. In some variations, a total amount oftherapeutic agent less than about 5.0 mg is administered subtenonally.In some variations, a total amount of therapeutic agent less than about4.5 mg is administered subtenonally. In some variations, a total amountof therapeutic agent less than about 4.0 mg is administeredsubtenonally. In some variations, a total amount of therapeutic agentless than about 3.5 mg is administered subtenonally. In some variations,a total amount of therapeutic agent less than about 3.0 mg isadministered subtenonally. In some variations, a total amount oftherapeutic agent less than about 2.5 mg is administered subtenonally.In some variations, a total amount of therapeutic agent less than about2 mg is administered subtenonally. In some variations, a total amount oftherapeutic agent less than about 1.2 mg is administered subtenonally.In some variations, a total amount of therapeutic agent less than about1.0 mg is administered subtenonally. In some variations, a total amountof therapeutic agent less than about 0.8 mg is administeredsubtenonally. In some variations, a total amount of therapeutic agentless than about 0.6 mg is administered subtenonally. In some variations,a total amount of therapeutic agent less than about 0.4 mg isadministered subtenonally. In some variations, a volume of a formulationis administered that contains an amount of therapeutic agent describedherein.

In some variations, a total amount of therapeutic agent less than about5 mg is administered subconjunctivally. In some variations, a totalamount of therapeutic agent less than about 5.0 mg is administeredsubconjunctivally. In some variations, a total amount of therapeuticagent less than about 4.5 mg is administered subconjunctivally. In somevariations, a total amount of therapeutic agent less than about 4.0 mgis administered subconjunctivally. In some variations, a total amount oftherapeutic agent less than about 3.5 mg is administeredsubconjunctivally. In some variations, a total amount of therapeuticagent less than about 3.0 mg is administered subconjunctivally. In somevariations, a total amount of therapeutic agent less than about 2.5 mgis administered subconjunctivally. In some variations, a total amount oftherapeutic agent less than about 2 mg is administeredsubconjunctivally. In some variations, a total amount of therapeuticagent less than about 1.2 mg is administered subconjunctivally. In somevariations, a total amount of therapeutic agent less than about 1.0 mgis administered subconjunctivally. In some variations, a total amount oftherapeutic agent less than about 0.8 mg is administeredsubconjunctivally. In some variations, a total amount of therapeuticagent less than about 0.6 mg is administered subconjunctivally. In somevariations, a total amount of therapeutic agent less than about 0.4 mgis administered subconjunctivally. In some variations, a volume of aformulation is administered that contains an amount of therapeutic agentdescribed herein.

In some variations, a total amount of therapeutic agent less than about200 μg is administered intravitreally. In some variations, a totalamount of therapeutic agent less than about 200 μg is administeredintravitreally. In some variations, a total amount of therapeutic agentless than about 300 μg is administered intravitreally. In somevariations, a total amount of therapeutic agent less than about 400 μgis administered intravitreally. In some variations, a total amount oftherapeutic agent less than about 500 μg is administered intravitreally.In some variations, a total amount of therapeutic agent less than about600 μg is administered intravitreally. In some variations, a totalamount of therapeutic agent less than about 800 μg is administeredintravitreally. In some variations, a total amount of therapeutic agentless than about 1 mg is administered intravitreally. In some variations,a total amount of therapeutic agent less than about 2 mg is administeredintravitreally. In some variations, a total amount of therapeutic agentless than about 2.5 mg is administered intravitreally. In somevariations, a total amount of therapeutic agent less than about 3 mg isadministered intravitreally. In some variations, a total amount oftherapeutic agent less than about 3.5 mg is administered intravitreally.In some variations, a total amount of therapeutic agent less than about4 mg is administered intravitreally. In some variations, a volume of aformulation is administered that contains an amount of therapeutic agentdescribed herein.

In some variations, a total amount of therapeutic agent administeredsubconjunctivally is any of between about 50 μg and about 3 mg, betweenabout 150 μg and about 750 μg, between about 300 μg and about 1000 μg,between about 300 μg and about 950 μg, between about 400 μg and about900 μg, between about 450 μg and about 850 μg, between about 500 μg andabout 800 μg, between about 550 μg and about 750 μg, or between about600 μg and about 700 μg. In some variations, a total amount oftherapeutic agent administered subconjunctivally is any of about 220 μg,about 440 μg, about 587 μg, about 630 μg, about 660 μg, about 880 μg,about 1320 μg, about 1760 μg, or about 2200 μg. In some variations, atotal amount of therapeutic agent administered subconjunctivally isabout 220 μg. In some variations, a total amount of therapeutic agentadministered subconjunctivally is about 440 μg. In some variations, atotal amount of therapeutic agent administered subconjunctivally isabout 660 μg. In some variations, a total amount of therapeutic agentadministered subconjunctivally is about 880 μg. In some variations, aliquid formulation containing an amount of therapeutic agent of 220 μgis subconjunctivally administered to a human subject by administeringabout 10 μl of a liquid formulation described herein. In somevariations, a liquid formulation containing an amount of therapeuticagent of 440 μg is subconjunctivally administered to a human subject byadministering about 20 μl of a liquid formulation described herein. Insome variations, a liquid formulation containing an amount oftherapeutic agent of 660 μg is subconjunctivally administered to a humansubject by administering about 30 μl of a liquid formulation describedherein. In some variations, a liquid formulation containing an amount oftherapeutic agent of 880 μg is subconjunctivally administered to a humansubject by administering about 40 μl of a liquid formulation describedherein.

In some variations, a total amount of therapeutic agent administeredintravitreally is any of between about 20 μg and about 750 μg, betweenabout 20 μg and about 500 μg, or between about 30 μg and about 200 μg.In some variations, a total amount of therapeutic agent administeredintravitreally is any of about 44 μg, about 110 μg, about 132 μg, about133.5 μg, about 176 μg, about 264 μg, about 352 μg, about 440 μg orabout 976 μg. In some variations, a total amount of therapeutic agentadministered intravitreally is about 44 μg. In some variations, a totalamount of therapeutic agent administered intravitreally is about 110 μg.In some variations, a total amount of therapeutic agent administeredintravitreally is about 967 μg. In some variations, a liquid formulationcontaining an amount of therapeutic agent of 44 μg is intravitreallyadministered to a human subject by administering about 2 μl of a liquidformulation described herein. In some variations, a liquid formulationcontaining an amount of therapeutic agent of 110 μg is intravitreallyadministered to a human subject by administering about 5 μl of a liquidformulation described herein. In some variations, a liquid formulationcontaining an amount of therapeutic agent of 176 μg is intravitreallyadministered to a human subject by administering about 8 μl of a liquidformulation described herein.

In some variations, a total volume of a liquid formulation describedherein is topically administered to a rabbit eye or a subject's,including but not limited to a human subject's eye that is less thanabout 1000 μl, less than about 900 μl, less than about 800 μl, less thanabout 700 μl, less than about 600 μl, less than about 500 μl, less thanabout 400 μl, less than about 300 μl, less than about 200 μl, less thanabout 150 μl, less than about 100 μl, less than about 90 μl, less thanabout 80 μl, less than about 70 μl, less than about 60 μl, less thanabout 50 μl, less than about 40 μl, less than about 30 μl, less thanabout 20 μl, less than about 10 μl, less than about 5 μl, less thanabout 3 μl, or less than about 1 μl. In some variations, a volume of aliquid formulation described herein is topically administered to arabbit eye or a subject's, including but not limited to a humansubject's eye that is less than about 200 μl. In some variations, avolume of a liquid formulation described herein is topicallyadministered to a rabbit eye or a subject's, including but not limitedto a human subject's eye that is less than about 100 μl. In somevariations, a volume of a liquid formulation described herein istopically administered to a rabbit eye or a subject's, including but notlimited to a human subject's eye that is between about 0.1 μl and about200 μl, between about 50 μl and about 200 μl, between about 200 μl andabout 300 μl, between about 300 μl and about 400 μl, between about 400μl and about 500 μl, between about 600 μl and about 700 μl, betweenabout 700 μl and about 800 μl, between about 800 μl and about 900 μl,between about 900 μl and about 1000 μl, between about 50 μl and about150 μl, between about 0.1 μl and about 100 μl, between about 0.1 μl andabout 50 μl, between about 1 μl and about 40 μl, between about 1 μl andabout 30 μl, between about 1 μl and about 20 μl, between about 1 μl andabout 10 μl, or between about 1 μl and about 5 μl. In some variations, atotal volume of a liquid formulation described herein is topicallyadministered to a rabbit eye or a subject's, including but not limitedto a human subject's eye that is between about 10 μl and about 200 μl.In some variations, a total volume of a liquid formulation describedherein is administered topically to a rabbit eye or a subject's,including but not limited to a human subject's eye that is between about0.1 μl and about 200 μl. In some variations, a total volume of a liquidformulation described herein is topically administered to a rabbit eyeor a subject's, including but not limited to a human subject's eye thatis between about 40 μl and about 160 μl. In some variations, a totalvolume of a liquid formulation described herein is topicallyadministered to a rabbit eye or a subject's, including but not limitedto a human subject's eye that is about 40 μl. In some variations, atotal volume of a liquid formulation described herein is topicallyadministered to a rabbit eye or a subject's, including but not limitedto a human subject's eye that is about 80 μl.

In some variations, a total amount of therapeutic agent less than about5 mg is administered topically. In some variations, a total amount oftherapeutic agent less than about 5.0 mg is administered topically. Insome variations, a total amount of therapeutic agent less than about 4.5mg is administered topically. In some variations, a total amount oftherapeutic agent less than about 4.0 mg is administered topically. Insome variations, a total amount of therapeutic agent less than about 3.5mg is administered topically. In some variations, a total amount oftherapeutic agent less than about 3.0 mg is administered topically. Insome variations, a total amount of therapeutic agent less than about 2.5mg is administered topically. In some variations, a total amount oftherapeutic agent less than about 2 mg is administered topically. Insome variations, a total amount of therapeutic agent less than about 1.2mg is administered topically. In some variations, a total amount oftherapeutic agent less than about 1.0 mg is administered topically. Insome variations, a total amount of therapeutic agent less than about 0.8mg is administered topically. In some variations, a total amount oftherapeutic agent less than about 0.6 mg is administered topically. Insome variations, a total amount of therapeutic agent less than about 0.4mg is administered topically. In some variations, a volume of aformulation is administered that contains an amount of therapeutic agentdescribed herein. In some variations, a total amount of therapeuticagent administered topically is any of between about 20 μg and about4000 μg, between about 10 μg and about 2000 μg, between about 10 μg and1750 μg, between about 1500 μg and 1000 μg, or between about 10 μg and1000 μg. In some variations, a total amount of therapeutic agentadministered topically is about 1660 μg. In some variations, a totalamount of therapeutic agent administered topically is about 880 μg. Insome variations, a total amount of therapeutic agent administeredtopically is 40 μg. In some variations, a total amount of therapeuticagent administered topically is about 28 μg.

In some variations, the therapeutic agent may be a limus compound. Insome variations, the therapeutic agent may be rapamycin or an analog orderivative thereof including those described in the Therapeutic Agentssection. In some variations, the therapeutic agent is rapamycin. In somevariations, the therapeutic agent is dasatinib.

“Total amount of a therapeutic agent” as used herein refers to the totalamount of a therapeutic agent administered during a singleadministration session by a patient and/or physician and/or othermedical professional. In some variations, a single administrationsession will involve a single administration of the therapeutic agent.In some variations, one administration session will include more thanone administration of the therapeutic agent. In some variations, oneadministration session will include a single route of administration. Insome variations, one administration session will include multiple routesof administration. Thus, in some variations, portions of the totalamount of the therapeutic agent are administered separately during asingle administration session. In such variations, the portions of thetotal amount that are administered separately may be administered by thesame and/or different routes of administration. In addition, in somevariations, portions of the total amount that are administeredseparately may be administered in the same and/or differentformulations.

“Total volume of a liquid formulation” as used herein refers to thetotal volume of a liquid formulation administered during a singleadministration session by a patient and/or physician and/or othermedical professional. In some variations, a single administrationsession will involve a single administration of the liquid formulation.In some variations, one administration session will include more thanone administration of the liquid formulation. In some variations, oneadministration session will include a single route of administration. Insome variations, one administration session will include multiple,different routes of administration. Thus, in some variations, portionsof the total volume are administered separately during a singleadministration session. In such variations, the portions of the totalvolume that are administered separately may be administered by the sameand/or by different routes of administration.

In some variations, the liquid formulation is administered in multipleocular locations. In some variations, the liquid formulation isadministered intravitreally and subconjunctivally. In some variations,the liquid formulation is administered intravitreally and subtenonally.In some variations, the liquid formulation is administered subtenonallyand subconjunctivally. In some variations, the liquid formulation isfirst administered intravitreally and at least one subsequentadministration is administered subconjunctivally. In some variations,the liquid formulation is first administered subconjunctivally and atleast one subsequent administration is administered intravitreally. Insome variations, the liquid formulation is first administeredintravitreally and at least one subsequent administration isadministered subtenonally. In some variations, the liquid formulation isfirst administered subconjunctivally and at least one subsequentadministration is administered subtenonally. In some variations, theliquid formulation is first administered subtenonally and at least onesubsequent administration is administered subconjunctivally. In somevariations, the liquid formulation is first administered subtenonallyand at least one subsequent administration is administeredintravitreally. In some variations, the liquid formulation is firstadministered intravitrealy, subconjunctivally, or subtenonally and atleast on subsequent administration is topically. In some variations, thefirst administration induces a drug response, and subsequentadministrations maintain the drug response. In some variations, the same(i.e., identical) liquid formulation is used for the different routes ofadministration, as a nonlimiting example for intravitreal andsubconjunctival administration. In some variations, administration tomultiple ocular locations occurs during one visit to the physician. Insome variations, administration to multiple ocular locations occursduring separate visits to the physician.

In some variations, a therapeutic agent is administered in multipleocular locations. In some variations, a therapeutic agent isadministered intravitreally and subconjunctivally. In some variations, atherapeutic agent is first administered intravitreally and at least onesubsequent administration is administered subconjunctivally. In somevariations, a therapeutic agent is first administered subconjunctivallyand at least one subsequent administration is administeredintravitreally. In some variations, a therapeutic agent is firstadministered intravitreally, subconjunctivally, or subtenonally and atleast on subsequent administration is topically. In some variations, thesame (i.e., identical) liquid formulation of the therapeutic agent isused for the first and subsequent administrations. In some variations, adifferent liquid formulation is used for intravitreal andsubconjunctival administration for the first and subsequentadministrations. In some variations, administration to multiple ocularlocations occurs during one visit to the physician. In some variations,administration to multiple ocular locations occurs during separatevisits to the physician.

In some variations the liquid formulations described herein areadministered in multiple subconjunctival locations within a period oftime, including without limitation within an hour of one another.Without being bound by theory, it is thought that such multipleadministrations, such as multiple injections, allow for a greater totaldose to be administered subconjunctivally than a single dose due to apotentially limited ability of the local ocular tissues to absorb largervolumes.

One liquid formulation described herein is an in situ gellingformulation. In situ gelling formulations, as described herein, comprisea therapeutic agent and a plurality of polymers which give a formulationthat forms a gel or a gel-like substance when placed in an aqueousmedium, including but not limited to an aqueous medium of the eye.

In some variations of the liquid formulations described herein, thetherapeutic agent is a solution or suspension of rapamycin in a liquidmedium. Liquid media include but are not limited to solvents, includingbut not limited to those in the Solubilization of Therapeutic Agentssection.

The liquid formulations described herein may comprise a solubilizingagent component. In some variations the solubilizing agent component isa surfactant. Note that there is some overlap between components thatmay be solvents and solubilizing agents, and therefore the samecomponent may in some systems be used as either a solvent or asolubilizing agent. A liquid formulation that comprises a therapeuticagent and a component that may be considered either a solvent or asolubilizing agent or surfactant will be considered a solvent if it isplaying the role of a solvent; if the component is not playing the roleof the solvent, the component may be considered a solubilizing agent orsurfactant.

Liquid formulations may optionally further comprise stabilizers,excipients, gelling agents, adjuvants, antioxidants, and/or othercomponents as described herein.

In some variations all components in the liquid formulation, other thanthe therapeutic agent, are liquid at room temperature.

In some variations, the liquid formulation comprises a release modifyingagent. In some variations, the release modifying agent is a film-formingpolymer component. The film-forming polymer component may comprise oneor more film-forming polymers. Any film-forming polymer may be used inthe excipient component. In some variations, the film-forming polymercomponent comprises a water insoluble film forming polymer. In somevariations, the release modifying agent component comprises an acrylicpolymer, including but not limited to polymethacrylate, including butnot limited to Eudragit RL.

Described herein are compositions and liquid formulations for deliveryof the therapeutic agents described in the Therapeutic Agents section.Delivery of therapeutic agents using the compositions and liquidformulations described herein may be used to treat, prevent, inhibit,delay the onset of, or cause the regression of the diseases andconditions described in the Diseases and Conditions section. Thecompositions and liquid formulations described herein may comprise anyof the therapeutic agents described in the Therapeutic Agents section,including but not limited to rapamycin. The compositions and liquidformulations described herein may comprise one or more than onetherapeutic agent. Other compositions and liquid formulations inaddition to those explicitly described herein may be used.

When the therapeutic agent is rapamycin, the compositions and liquidformulations may be used to maintain an amount of rapamycin in thevitreous effective to treat wet AMD. In one nonlimiting example, it isbelieved that a liquid formulation delivering rapamycin to maintain aconcentration of rapamycin of about 10 pg/ml to about 2 μg/ml in thevitreous over a period of time may be used for the treatment of wet AMD.When the rapamycin is in a liquid formulation that forms a non-dispersedmass, the stated concentration of rapamycin represents the amount thatis effectively treating the disease or condition of the eye, and notmerely present in the form of the non-dispersed mass. In anothernonlimiting example, it is believed that a delivery system deliveringrapamycin to maintain a concentration of rapamycin of about 0.01 μg/mgto about 10 ng/mg in the retina choroid tissues over a period of timemay be used for treatment of wet AMD. Other therapeutically effectiveamounts of therapeutic agent are also possible, and can be readilydetermined by one of skill in the art given the teachings herein.

When the therapeutic agent is rapamycin, the compositions and liquidformulations described herein may be used to deliver a dose of rapamycinto a subject, including but not limited to a human subject or to the eyeof a subject. In one nonlimiting example, it is believed that a liquidformulation containing a dose of about 20 μg to about 4 mg may be usedfor the treatment of wet AMD.

In some variations the therapeutic agent in the liquid formulationcomprises between about 0.01 to about 30% of the total weight of thecomposition; between about 0.05 to about 15%; between about 0.1 to about10%; between about 1 to about 5%; or between about 5 to about 15%;between about 8 to about 10%; between about 0.01 to about 1%; betweenabout 0.05 to about 5%; between about 0.1 to about 0.2%; between about0.2 to about 0.3%; between about 0.3 to about 0.4%; between about 0.4 toabout 0.5%; between about 0.5 to about 0.6%; between about 0.6 to about0.7%; between about 0.7 to about 1%; between about 1 to about 5%;between about 5 to about 10%; between about 15 to about 30%, betweenabout 20 to about 30%; or between about 25 to about 30%.

In some variations the therapeutic agent in the liquid formulationcomprises between about 0.001 to about 1.00% of the total weight of thecomposition. In some variations the therapeutic agent in the liquidformulation comprises any of about 0.07%, about 0.08%, 0.09%, 0.17%,1.38%, 1.47%, 2%, 4%, 4.84%, or 5% of the total weight of thecomposition. In some variations, the therapeutic agent may be a limuscompound. In some variations, the therapeutic agent may be rapamycin oran analog or derivative thereof including those described in theTherapeutic Agents section. In some variations, the therapeutic agent israpamycin. In some variations, the therapeutic agent is dasatinib.

Those of skill in the art, based on the teachings herein can determinewhat amount or concentration of a given therapeutic agent is equivalentto an amount or concentration of rapamycin by, for example,administering the therapeutic agent at various amounts or concentrationsto a disease model system, such as an in vivo or in vivo model system,and comparing the results in the model system relative to the results ofvarious amounts or concentrations of rapamycin. Those of skill in theart, based on the teachings herein can also determine what amount orconcentration of a given therapeutic agent is equivalent to an amount orconcentration of rapamycin by reviewing the scientific literature forexperiments performed comparing rapamycin to other therapeutic agents.It is understood that even the same therapeutic agent may have adifferent equivalent level of rapamycin when, for example, a differentdisease or disorder is being evaluated, or a different type offormulation is used. Nonlimiting examples of scientific references withcomparative studies of rapamycin and other therapeutic agents on oculardisease are Ohia et al., Effects of steroids and immunosuppressive drugson endotoxin-uveitis in rabbits, J. Ocul. Pharmacol. 8(4):295-307(1992); Kulkami, Steroidal and nonsteroidal drugs in endotoxin-induceduveitis, J. Ocul. Pharmacol. 10(1):329-34 (1994); Hafizi et al.,Differential effects of rapamycin, cyclosporine A, and FK506 on humancoronary artery smooth muscle cell proliferation and signaling, VasculPharmacol. 41(4-5):167-76 (2004); and US 2005/0187241.

For example, in a model for wet AMD, if a therapeutic agent is found tobe approximately 10-fold less potent or efficacious than rapamycin inthe treatment of wet AMD, a concentration of 10 ng/ml of the therapeuticagent would be equivalent to a 1 ng/ml concentration of rapamycin. Or ifa therapeutic agent is found to be approximately 10-fold less potent orefficacious than rapamycin in the treatment of wet AMD, a 10-fold amountof the therapeutic agent would be administered relative to the amount ofrapamycin.

The solvent component may comprise, for instance, between about 0.01 toabout 99.9% of the total weight of the composition; between about 0.1 toabout 99%; between about 25 to about 55%; between about 30 to about 50%;or between about 35 to about 45%; between about 0.1 to about 10%;between about 10 to about 20%; between about 20 to about 30%; betweenabout 30 to about 40%; between about 40 to about 45%; between about 40to about 45%; between about 45 to about 50%; between about 50 to about60%; between about 50 to about 70%; between about 70 to about 80%;between about 80 to about 90%; or between about 90 to about 100%.

The solubilizing agent component may comprise, for instance, betweenabout 0.01 to about 30% of the total weight of the composition; betweenabout 0.1 to about 20%; between about 2.5 to about 15%; between about 10to about 15%; or between about 5 to about 10%; between about 8 to about12%; between about 10 to about 20%; between about 20 to about 30%.

In some variations, the liquid formulations described herein have aviscosity of between 40% and 120% centipoise. In some variations theliquid formulations described herein have a viscosity of between 60% and80% centipoise.

In some variations the liquid formulations described herein comprise atherapeutic agent and a solvent component. The solvent component maycomprise a single solvent or a combination of solvents. The therapeuticagent component may comprise a single therapeutic agent or a combinationof therapeutic agents. In some variations, the solvent is glycerin,dimethylsulfoxide, N-methylpyrrolidone, dimethyl acetamide (DMA),dimethyl formamide, glycerol formal, ethoxy diglycol, triethylene glycoldimethyl ether, triacetin, diacetin, corn oil, acetyl triethyl citrate(ATC), ethyl lactate, polyglycolated capryl glyceride, γ butyrolactone,dimethyl isosorbide, benzyl alcohol, ethanol, isopropyl alcohol,polyethylene glycol of various molecular weights, including but notlimited to PEG 300 and PEG 400, or propylene glycol, or a mixture of oneor more thereof.

In some variations the liquid formulations described herein aresolutions, and comprise a therapeutic agent and a solvent component. Insome variations the solvent component comprises ethanol. In somevariations the solvent component comprises ethanol and a polyethyleneglycol, including but not limited to a liquid polyethylene glycol,including but not limited to one or more of PEG 300 or PEG 400.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of polyethylene glycol. In some variations theliquid formulations described herein contain no greater than about 250μl, no greater than about 200 μl, no greater than about 150 μl, nogreater than about 125 μl, no greater than about 100 μl, no greater thanabout 75 μl, no greater than about 50 μl, no greater than about 25 μl,no greater than about 20 μl, no greater than about 15 μl, no greaterthan about 10 μl, no greater than about 7.5 μl, no greater than about 5μl, no greater than about 2.5 μl, no greater than about 1.0 μl, or nogreater than about 0.5 μl of polyethylene glycol. Formulationscontaining polyethylene glycol may contain, for example, PEG 300 or PEG400.

In some variations, the liquid formulations described herein aresuspensions, and comprise a therapeutic agent and a diluent component.In some variations, the diluent component comprises one or morecomponents listed herein as solvents or solubilizing agents, wherein theresulting mixture is a suspension.

In some variations the liquid formulation is partly a solution andpartly a suspension.

In some variations the liquid formulation is an in situ gellingformulation, and comprises a therapeutic agent and a polymer component,wherein the polymer component may comprise a plurality of polymers. Insome variations, the liquid formulation comprises a polymethacrylatepolymer. In some variations, the liquid formulation comprises apolyvinylpyrrolidone polymer.

Some variations of liquid formulations include a therapeutic agent oragents such as but not limited to rapamycin between about 0.01% andabout 20% by weight of the total, a solvent between about 5% and about15% by weight of the total, a solubilizing agent including but notlimited to a surfactant between about 5% and about 15% by weight of thetotal, with water as the primary remaining component. In some variationsthe formulations further comprise stabilizing agents, excipients,adjuvants, or antioxidants, between about 0 and about 40% by weight ofthe total.

In some variations, a liquid formulation comprises up to about 5%therapeutic agent, including but not limited to rapamycin, per weight ofthe total; and up to about 99.9% of a solvent component, by weight ofthe total. In some variations the liquid formulation comprises up toabout 5% therapeutic agent, including but not limited to rapamycin, perweight of the total; and up to about 99.9% of a diluent component.

In some variations, a liquid formulation may comprise up to about 5%therapeutic agent, including but not limited to rapamycin, per weight ofthe total; up to about 10% solvent by weight of the total; and up toabout 85% of a solubilizing component, by weight of the total. In somevariations the solubilizing component is an aqueous solution of asurfactant.

A plurality of polymers component may comprise, for instance, betweenabout 0.01 to about 30% of the total weight of the composition; betweenabout 0.1 to about 20%; between about 2.5 to about 15%; between about 10to about 15%; between about 3 to about 5%; between about 5 to about 10%;between about 8 to about 12%; between about 10 to about 20%; or betweenabout 20 to about 30%.

Some variations of liquid formulations includes a therapeutic agent oragents such as but not limited to rapamycin between about 0.01% andabout 20% by weight of the total, a solvent component between about 60%and about 98% by weight of the total, and a plurality of polymers, whosecombined percentage is between about 0.1% and about 15% by weight of thetotal. In some variations the formulations further comprise stabilizingagents, excipients, adjuvants, or antioxidants, between about 0 andabout 40% by weight of the total.

In some variations, a liquid formulation may comprise about 4%therapeutic agent, including but not limited to rapamycin, per weight ofthe total; about 91% solvent by weight of the total; and about 5%polymeric component, per weight of the total.

In some variations, the liquid formulation comprises about 2% (w/w)rapamycin, about 4% (w/w) ethanol, and about 94% (w/w) PEG 400. In somevariations, liquid formulation is administered by subconjunctival,subtenon, and/or intravitreal injection. In some variations, the liquidformulation is used to treat or prevent an ocular disease. In somevariations, the ocular disease is wet AMD. In some variations, theocular disease is dry AMD. In some variations, the ocular disease ismacular edema.

In some variations, the liquid formulation comprises about 1.69% (w/w)rapamycin, about 3.39% (w/w) ethanol, about 79.54% (w/w) PEG 400, andabout 15.38% (w/w) aqueous liquid (e.g., water or saline). In somevariations, liquid formulation is administered by subconjunctival,subtenon, and/or intravitreal injection. In some variations, the liquidformulation is used to treat or prevent an ocular disease. In somevariations, the ocular disease is wet AMD. In some variations, theocular disease is dry AMD. In some variations, the ocular disease ismacular edema.

In some variations, the liquid formulation comprises about 1.47% (w/w)rapamycin, about 2.93% (w/w) ethanol, about 94% (w/w) PEG 400, and about26.6% (w/w) aqueous liquid (e.g., water or saline). In some variations,liquid formulation is administered by subconjunctival, subtenon, and/orintravitreal injection. In some variations, the liquid formulation isused to treat or prevent an ocular disease. In some variations, theocular disease is wet AMD. In some variations, the ocular disease is dryAMD. In some variations, the ocular disease is macular edema.

In some variations, the liquid formulation comprises about 1.38% (w/w)rapamycin, about 2.75% (w/w) ethanol, about 64.62% (w/w) PEG 400, andabout 31.25% (w/w) aqueous liquid (e.g., water or saline). In somevariations, liquid formulation is administered by subconjunctival,subtenon, and/or intravitreal injection. In some variations, the liquidformulation is used to treat or prevent an ocular disease. In somevariations, the ocular disease is wet AMD. In some variations, theocular disease is dry AMD. In some variations, the ocular disease ismacular edema.

In some variations the liquid formulations described herein comprise atherapeutic agent and a solvent component. In some variations, thesolvent component may comprise a solvent having a hygroscopicity that isequal to or greater than that of polyethylene glycol 400 (PEG 400). Insome variations, the solvent component may comprise a solvent having ahygroscopicity that is greater than that of polyethylene glycol 400 (PEG400). In some variations, the solvent component may comprise a solventhaving a hygroscopicity that is about equal to that of polyethyleneglycol 400 (PEG 400). In some variations, the solvent component maycomprise a solvent having a hygroscopicity that is less than that ofpolyethylene glycol 400 (PEG 400). In some variations, the liquidformulation is about as hygroscopic as a liquid formulation consistingof 2% rapamycin, 4% ethanol, and 94% PEG 400. In some variations, theliquid formulation is less hygroscopic as a liquid formulationconsisting of 2% rapamycin, 4% ethanol, and 94% PEG 400.

In some variations, parameters are used to measure hygroscopicity of amaterial include, but are not limited to, critical relative humidity(which may be measured by “moisture meters” (e.g., Hygrometers (i.e.,Thermo Hygrometer Model 625, manufactured by BK Precision))),hygroscopicity potential, hygroscopicity coefficient, or heats ofabsorption (which may be measured by a Differential Scanning Calorimetry(DSC). Those of ordinary skill in the art will find it routine toidentify solvents having a hygroscopicity that is about equal to, lessthan, or greater than that of polyethylene glycol 400 (PEG 400) giventhe teachings herein.

The solvent component may comprise a single solvent or a combination ofsolvents. The therapeutic agent component may comprise a singletherapeutic agent or a combination of therapeutic agents. In somevariations, the solvent is N-methylpyrrolidone (NMP), dimethyl-acetamine(DMA), dimethyl sulfoxide (DMSO), propylene glycol (PG), polyethyleneglycol 600 (PEG 600), polyethylene glycol 400 (PEG 400), ethanol, or amixture of one or more thereof. In some variations, the solventcomponent comprises a combination of solvents includingN-methylpyrrolidone (NMP), dimethyl-acetamine (DMA), or dimethylsulfoxide (DMSO). In some variations, the solvent component comprises acombination of solvents including propylene glycol (PG), polyethyleneglycol 600 (PEG 600), or polyethylene glycol 400 (PEG 400). In somevariation, the solvent component may comprise a combination of at leasttwo solvents. In some variations, the at least two solvents comprising afirst solvent such as N-methylpyrrolidone (NMP), dimethyl-acetamine(DMA), or dimethyl sulfoxide (DMSO) and a second solvent such aspropylene glycol (PG), polyethylene glycol 600 (PEG 600), orpolyethylene glycol 400 (PEG 400). In some variations, the solventcomponent may comprise N-methylpyrrolidone (NMP) and propylene glycol(PG). In some variations, the solvent component may compriseN-methylpyrrolidone (NMP) and polyethylene glycol 600 (PEG 600). In somevariations, the solvent component may comprise dimethyl-acetamine (DMA)and propylene glycol (PG). In some variations, the solvent component maycomprise dimethyl-acetamine (DMA) and polyethylene glycol 400 (PEG 400).In some variations, the solvent component may comprise dimethylsulfoxide (DMSO) and propylene glycol (PG). In some variations, thesolvent component may comprise N-methylpyrrolidone (NMP) andpolyethylene glycol 400 (PEG 400). In some variations, the solventcomponent may further comprise ethanol. In some variations, the solventcomponent may further comprise water. In some variations, the solventcomponent may have a hygroscopicity that is about equal to, less than,or greater than that of polyethylene glycol 400 (PEG 400). In somevariations, liquid formulation is administered by subconjunctival,subtenon, and/or intravitreal injection. In some variations, the liquidformulation is used to treat or prevent an ocular disease. In somevariations, the ocular disease is wet AMD. In some variations, theocular disease is dry AMD. In some variations, the ocular disease ismacular edema.

In some variations, the solvent is polyethoxylated castor oil (e.g.,Cremophor (PEG 35 castor oil)), monoglycerides and/or diglycerides ofcaprylic acid (e.g., Capmul MCM (C8)), nonionic polymer of the alkylaryl polyether alcohol (e.g., tyloxapol (ethoxylated p-tert-octylphenolformaldehyde polymer)), 50% phosphatidylcholine in propyleneglycol/ethanol carrier (e.g., Phosal® 50PG), propylene glycolmonolaurate, propylene glycol dicaprylate/dicaprate, macrogol 15hydroxystearate, ethanol, or a mixture of one or more thereof. In somevariation, the solvent may comprise a combination of at least twosolvents. In some variations, the at least two solvents comprising afirst solvent such as polyethoxylated castor oil (e.g., Cremophor (PEG35 castor oil)), propylene glycol monolaurate, propylene glycoldicaprylate/dicaprate, macrogol 15 hydroxystearate, or nonionic polymerof the alkyl aryl polyether alcohol (e.g., tyloxapol (ethoxylatedp-tert-octylphenol formaldehyde polymer)) and a second solvent such asmonoglycerides and/or diglycerides of caprylic acid (e.g., Capmul MCM(C8)). In some variations, the solvent may further comprise ethanol. Insome variations, the solvent may further comprise water. In somevariations, liquid formulation is topically administered. In somevariations, the liquid formulation is administered as eye drops. In somevariations, the liquid formulation is used to treat or prevent an oculardisease. In some variations, the ocular disease is wet AMD. In somevariations, the ocular disease is dry AMD. In some variations, theocular disease is macular edema.

The solvent component may comprise water, for instance, between about0.01 to about 99.9% of the total weight of the composition; betweenabout 0.1 to about 99%; between about 25 to about 55%; between about 30to about 50%; or between about 35 to about 45%; between about 0.1 toabout 10%; between about 10 to about 20%; between about 20 to about 30%;between about 30 to about 40%; between about 40 to about 45%; betweenabout 40 to about 45%; between about 45 to about 50%; between about 50to about 60%; between about 50 to about 70%; between about 70 to about80%; between about 80 to about 90%; or between about 90 to about 100%.In some variations, water comprises between about 15 to about 30% (w/w)of the liquid formulation. In some variations, water comprises betweenabout 80 to about 90% (w/w) of the liquid formulation. In somevariations, water comprises at least 15% (w/w). In some variations,water comprises at least 20% (w/w) of the liquid formulation. In somevariations, water comprises at least 25% (w/w) of the liquidformulation. In some variations, water comprises at least about 5percent (w/w) of the liquid formulation. In some variations, watercomprises about 16% (w/w) of the liquid formulation. In some variations,water comprises about 22% (w/w). In some variations, water comprisesabout 28% (w/w) of the liquid formulation. In some variations, watercomprises about 83% (w/w) of the liquid formulation.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of water. In some variations the liquidformulations described herein contain no greater than about 250 μl, nogreater than about 200 μl, no greater than about 150 μl, no greater thanabout 125 μl, no greater than about 100 μl, no greater than about 75 μl,no greater than about 50 μl, no greater than about 25 μl, no greaterthan about 20 μl, no greater than about 15 μl, no greater than about 10μl, no greater than about 7.5 μl, no greater than about 5 μl, no greaterthan about 2.5 μl, no greater than about 1.0 μl, or no greater thanabout 0.5 μl of water.

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of water.

The solvent component may comprise N-methylpyrrolidone (NMP), forinstance, between about 0.01 to about 99.9% of the total weight of thecomposition; between about 0.1 to about 99%; between about 25 to about55%; between about 30 to about 50%; or between about 35 to about 45%;between about 0.1 to about 10%; between about 10 to about 20%; betweenabout 20 to about 30%; between about 30 to about 40%; between about 40to about 45%; between about 40 to about 45%; between about 45 to about50%; between about 50 to about 60%; between about 50 to about 70%;between about 70 to about 80%; between about 80 to about 90%; or betweenabout 90 to about 100%. In some variations, NMP comprises between about10 to about 40% (w/w) of the liquid formulation. In some variations, NMPcomprises at least 10% (w/w) of the liquid formulation. In somevariations, NMP comprises at least 11% (w/w) of the liquid formulation.In some variations, NMP comprises at least 25% (w/w) of the liquidformulation. In some variations, NMP comprises at least 35% (w/w) of theliquid formulation. In some variations, NMP comprises about 10% (w/w) ofthe liquid formulation. In some variations, NMP comprises about 28%(w/w) of the liquid formulation. In some variations, NMP comprises about39% (w/w) of the liquid formulation.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of N-methylpyrrolidone (NMP). In somevariations the liquid formulations described herein contain no greaterthan about 250 μl, no greater than about 200 μl, no greater than about150 μl, no greater than about 125 μl, no greater than about 100 μl, nogreater than about 75 μl, no greater than about 50 μl, no greater thanabout 25 μl, no greater than about 20 μl, no greater than about 15 μl,no greater than about 10 μl, no greater than about 7.5 μl, no greaterthan about 5 μl, no greater than about 2.5 μl, no greater than about 1.0μl, or no greater than about 0.5 μl of N-methylpyrrolidone (NMP).

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of N-methylpyrrolidone(NMP).

The solvent component may comprise dimethyl-acetamine (DMA), forinstance, between about 0.01 to about 99.9% of the total weight of thecomposition; between about 0.1 to about 99%; between about 25 to about55%; between about 30 to about 50%; or between about 35 to about 45%;between about 0.1 to about 10%; between about 10 to about 20%; betweenabout 20 to about 30%; between about 30 to about 40%; between about 40to about 45%; between about 40 to about 45%; between about 45 to about50%; between about 50 to about 60%; between about 50 to about 70%;between about 70 to about 80%; between about 80 to about 90%; or betweenabout 90 to about 100%. In some variations, DMA comprises between about10 to about 40% (w/w) of the liquid formulation. In some variations, DMAcomprises at least 10% (w/w) of the liquid formulation. In somevariations, DMA comprises about 15% (w/w) of the liquid formulation. Insome variations, DMA comprises about 11% (w/w) of the liquidformulation. In some variations, DMA comprises about 10% (w/w) of theliquid formulation.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of dimethyl-acetamine (DMA). In somevariations the liquid formulations described herein contain no greaterthan about 250 μl, no greater than about 200 μl, no greater than about150 μl, no greater than about 125 μl, no greater than about 100 μl, nogreater than about 75 μl, no greater than about 50 μl, no greater thanabout 25 μl, no greater than about 20 μl, no greater than about 15 μl,no greater than about 10 μl, no greater than about 7.5 μl, no greaterthan about 5 μl, no greater than about 2.5 μl, no greater than about 1.0μl, or no greater than about 0.5 μl of dimethyl-acetamine (DMA).

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of dimethyl-acetamine(DMA).

The solvent component may comprise dimethyl sulfoxide (DMSO), forinstance, between about 0.01 to about 99.9% of the total weight of thecomposition; between about 0.1 to about 99%; between about 25 to about55%; between about 30 to about 50%; or between about 35 to about 45%;between about 0.1 to about 10%; between about 10 to about 20%; betweenabout 20 to about 30%; between about 30 to about 40%; between about 40to about 45%; between about 40 to about 45%; between about 45 to about50%; between about 50 to about 60%; between about 50 to about 70%;between about 70 to about 80%; between about 80 to about 90%; or betweenabout 90 to about 100%. In some variations, DMSO comprises between about10 to about 40% (w/w) of the liquid formulation. In some variations,DMSO comprises at least 10% (w/w) of the liquid formulation. In somevariations, DMSO comprises at least 11% (w/w) of the liquid formulation.In some variations, DMSO comprises about 10% (w/w) of the liquidformulation.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of dimethyl sulfoxide (DMSO). In somevariations the liquid formulations described herein contain no greaterthan about 250 μl, no greater than about 200 μl, no greater than about150 μl, no greater than about 125 μl, no greater than about 100 μl, nogreater than about 75 μl, no greater than about 50 μl, no greater thanabout 25 μl, no greater than about 20 μl, no greater than about 15 μl,no greater than about 10 μl, no greater than about 7.5 μl, no greaterthan about 5 μl, no greater than about 2.5 μl, no greater than about 1.0μl, or no greater than about 0.5 μl of dimethyl sulfoxide (DMSO).

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of dimethyl sulfoxide(DMSO).

The solvent component may comprise propylene glycol (PG), for instance,between about 0.01 to about 99.9% of the total weight of thecomposition; between about 0.1 to about 99%; between about 25 to about55%; between about 30 to about 50%; or between about 35 to about 45%;between about 0.1 to about 10%; between about 10 to about 20%; betweenabout 20 to about 30%; between about 30 to about 40%; between about 40to about 45%; between about 40 to about 45%; between about 45 to about50%; between about 50 to about 60%; between about 50 to about 70%;between about 70 to about 80%; between about 80 to about 90%; or betweenabout 90 to about 100%. In some variations, PG comprises between about35 to about 75% (w/w) of the liquid formulation. In some variations, PGcomprises at least 35% (w/w) of the liquid formulation. In somevariations, PG comprises at least 70% (w/w) of the liquid formulation.In some variations, PG comprises less than about 90% (w/w) of the liquidformulation. In some variations, PG comprises about 39% (w/w) of theliquid formulation. In some variations, PG comprises about 74% (w/w) ofthe liquid formulation.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of propylene glycol (PG). In some variationsthe liquid formulations described herein contain no greater than about250 μl, no greater than about 200 μl, no greater than about 150 μl, nogreater than about 125 μl, no greater than about 100 μl, no greater thanabout 75 μl, no greater than about 50 μl, no greater than about 25 μl,no greater than about 20 μl, no greater than about 15 μl, no greaterthan about 10 μl, no greater than about 7.5 μl, no greater than about 5μl, no greater than about 2.5 μl, no greater than about 1.0 μl, or nogreater than about 0.5 μl of propylene glycol (PG).

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of propylene glycol (PG).

The solvent component may comprise polyethylene glycol (PEG), forinstance, between about 0.01 to about 99.9% of the total weight of thecomposition; between about 0.1 to about 99%; between about 25 to about55%; between about 30 to about 50%; or between about 35 to about 45%;between about 0.1 to about 10%; between about 10 to about 20%; betweenabout 20 to about 30%; between about 30 to about 40%; between about 40to about 45%; between about 40 to about 45%; between about 45 to about50%; between about 50 to about 60%; between about 50 to about 70%;between about 70 to about 80%; between about 80 to about 90%; or betweenabout 90 to about 100%. In some variations, PEG comprises between about35 to about 75% (w/w) of the liquid formulation. In some variations, PEGcomprises at least 35% (w/w) of the liquid formulation. In somevariations, PEG comprises at least 70% (w/w) of the liquid formulation.In some variations, PEG comprises less than about 90% (w/w) of theliquid formulation. In some variations, PEG comprises about 39% (w/w) ofthe liquid formulation. In some variations, PEG comprises about 74%(w/w) of the liquid formulation. In some variations, PEG comprises about80% (w/w) of the liquid formulation. In some variations, the PEG ispolyethylene glycol 400. In some variations, the PEG is polyethyleneglycol 600.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of polyethylene glycol. In some variations theliquid formulations described herein contain no greater than about 250μl, no greater than about 200 μl, no greater than about 150 μl, nogreater than about 125 μl, no greater than about 100 μl, no greater thanabout 75 μl, no greater than about 50 μl, no greater than about 25 μl,no greater than about 20 μl, no greater than about 15 μl, no greaterthan about 10 μl, no greater than about 7.5 μl, no greater than about 5μl, no greater than about 2.5 μl, no greater than about 1.0 μl, or nogreater than about 0.5 μl of polyethylene glycol. In some variations,the PEG is polyethylene glycol 400. In some variations, the PEG ispolyethylene glycol 600.

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of polyethylene glycol. Insome variations, the PEG is polyethylene glycol 400. In some variations,the PEG is polyethylene glycol 600.

The solvent component may comprise ethanol, for instance, between about0.01 to about 99.9% of the total weight of the composition; betweenabout 0.1 to about 99%; between about 25 to about 55%; between about 30to about 50%; or between about 35 to about 45%; between about 0.1 toabout 10%; between about 10 to about 20%; between about 20 to about 30%;between about 30 to about 40%; between about 40 to about 45%; betweenabout 40 to about 45%; between about 45 to about 50%; between about 50to about 60%; between about 50 to about 70%; between about 70 to about80%; between about 80 to about 90%; or between about 90 to about 100%.In some variations, ethanol comprises less than about 10% (w/w) of theliquid formulation. In some variations, ethanol comprises less thanabout 5% (w/w) of the liquid formulation. In some variations, ethanolcomprises less than about 1% (w/w) of the liquid formulation. In somevariations, ethanol comprises less than about 0.5% (w/w) of the liquidformulation. In some variations, ethanol comprises about 5% (w/w) of theliquid formulation. In some variations, ethanol comprises about 2.9%(w/w) of the liquid formulation. In some variations, ethanol comprisesabout 0.35% (w/w) of the liquid formulation.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of ethanol. In some variations the liquidformulations described herein contain no greater than about 250 μl, nogreater than about 200 μl, no greater than about 150 μl, no greater thanabout 125 μl, no greater than about 100 μl, no greater than about 75 μl,no greater than about 50 μl, no greater than about 25 μl, no greaterthan about 20 μl, no greater than about 15 μl, no greater than about 10μl, no greater than about 7.5 μl, no greater than about 5 μl, no greaterthan about 2.5 μl, no greater than about 1.0 μl, or no greater thanabout 0.5 μl of ethanol.

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of ethanol.

The solvent component may comprise polyethoxylated castor oil (e.g.,Cremophor (PEG 35 castor oil)), for instance, between about 0.01 toabout 99.9% of the total weight of the composition; between about 0.1 toabout 99%; between about 25 to about 55%; between about 30 to about 50%;or between about 35 to about 45%; between about 0.1 to about 10%;between about 10 to about 20%; between about 20 to about 30%; betweenabout 30 to about 40%; between about 40 to about 45%; between about 40to about 45%; between about 45 to about 50%; between about 50 to about60%; between about 50 to about 70%; between about 70 to about 80%;between about 80 to about 90%; or between about 90 to about 100%. Insome variations, polyethoxylated castor oil (e.g., Cremophor (PEG 35castor oil)) comprises less than about 10% (w/w) of the liquidformulation. In some variations, polyethoxylated castor oil (e.g.,Cremophor (PEG 35 castor oil)) comprises about 8.35% (w/w) of the liquidformulation. In some variations, polyethoxylated castor oil (e.g.,Cremophor (PEG 35 castor oil)) comprises about 7.18% (w/w) of the liquidformulation.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of polyethoxylated castor oil (e.g., Cremophor(PEG 35 castor oil)). In some variations the liquid formulationsdescribed herein contain no greater than about 250 μl, no greater thanabout 200 μl, no greater than about 150 μl, no greater than about 125μl, no greater than about 100 μl, no greater than about 75 μl, nogreater than about 50 μl, no greater than about 25 μl, no greater thanabout 20 μl, no greater than about 15 μl, no greater than about 10 μl,no greater than about 7.5 μl, no greater than about 5 μl, no greaterthan about 2.5 μl, no greater than about 1.0 μl, or no greater thanabout 0.5 μl of polyethoxylated castor oil (e.g., Cremophor (PEG 35castor oil)).

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of polyethoxylated castoroil (e.g., Cremophor (PEG 35 castor oil)).

The solvent component may comprise monoglycerides and/or diglycerides ofcaprylic acid (e.g., Capmul MCM (C8)), for instance, between about 0.01to about 99.9% of the total weight of the composition; between about 0.1to about 99%; between about 25 to about 55%; between about 30 to about50%; or between about 35 to about 45%; between about 0.1 to about 10%;between about 10 to about 20%; between about 20 to about 30%; betweenabout 30 to about 40%; between about 40 to about 45%; between about 40to about 45%; between about 45 to about 50%; between about 50 to about60%; between about 50 to about 70%; between about 70 to about 80%;between about 80 to about 90%; or between about 90 to about 100%. Insome variations, monoglycerides and/or diglycerides of caprylic acid(e.g., Capmul MCM (C8)) comprises less than about 10% (w/w) of theliquid formulation. In some variations, monoglycerides and/ordiglycerides of caprylic acid (e.g., Capmul MCM (C8)) comprises about9.47% (w/w) of the liquid formulation. In some variations,monoglycerides and/or diglycerides of caprylic acid (e.g., Capmul MCM(C8)) comprises about 8.37% (w/w) of the liquid formulation. In somevariations, monoglycerides and/or diglycerides of caprylic acid (e.g.,Capmul MCM (C8)) comprises about 8.15% (w/w) of the liquid formulation.In some variations, monoglycerides and/or diglycerides of caprylic acid(e.g., Capmul MCM (C8)) comprises about 6.81% (w/w) of the liquidformulation.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of monoglycerides and/or diglycerides ofcaprylic acid (e.g., Capmul MCM (C8)). In some variations the liquidformulations described herein contain no greater than about 250 μl, nogreater than about 200 μl, no greater than about 150 μl, no greater thanabout 125 μl, no greater than about 100 μl, no greater than about 75 μl,no greater than about 50 μl, no greater than about 25 μl, no greaterthan about 20 μl, no greater than about 15 μl, no greater than about 10μl, no greater than about 7.5 μl, no greater than about 5 μl, no greaterthan about 2.5 μl, no greater than about 1.0 μl, or no greater thanabout 0.5 μl of monoglycerides and/or diglycerides of caprylic acid(e.g., Capmul MCM (C8)).

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of monoglycerides and/ordiglycerides of caprylic acid (e.g., Capmul MCM (C8)).

The solvent component may comprise nonionic polymer of the alkyl arylpolyether alcohol (e.g., tyloxapol (ethoxylated p-tert-octylphenolformaldehyde polymer)), for instance, between about 0.01 to about 99.9%of the total weight of the composition; between about 0.1 to about 99%;between about 25 to about 55%; between about 30 to about 50%; or betweenabout 35 to about 45%; between about 0.1 to about 10%; between about 10to about 20%; between about 20 to about 30%; between about 30 to about40%; between about 40 to about 45%; between about 40 to about 45%;between about 45 to about 50%; between about 50 to about 60%; betweenabout 50 to about 70%; between about 70 to about 80%; between about 80to about 90%; or between about 90 to about 100%. In some variations,nonionic polymer of the alkyl aryl polyether alcohol (e.g., tyloxapol(ethoxylated p-tert-octylphenol formaldehyde polymer)) comprises lessthan about 10% (w/w) of the liquid formulation. In some variations,nonionic polymer of the alkyl aryl polyether alcohol (e.g., tyloxapol(ethoxylated p-tert-octylphenol formaldehyde polymer)) comprises about8.12% (w/w) of the liquid formulation. In some variations, nonionicpolymer of the alkyl aryl polyether alcohol (e.g., tyloxapol(ethoxylated p-tert-octylphenol formaldehyde polymer)) comprises about6.81% (w/w) of the liquid formulation.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of nonionic polymer of the alkyl arylpolyether alcohol (e.g., tyloxapol (ethoxylated p-tert-octylphenolformaldehyde polymer)). In some variations the liquid formulationsdescribed herein contain no greater than about 250 μl, no greater thanabout 200 μl, no greater than about 150 μl, no greater than about 125μl, no greater than about 100 μl, no greater than about 75 μl, nogreater than about 50 μl, no greater than about 25 μl, no greater thanabout 20 μl, no greater than about 15 μl, no greater than about 10 μl,no greater than about 7.5 μl, no greater than about 5 μl, no greaterthan about 2.5 μl, no greater than about 1.0 μl, or no greater thanabout 0.5 μl of nonionic polymer of the alkyl aryl polyether alcohol(e.g., tyloxapol (ethoxylated p-tert-octylphenol formaldehyde polymer)).

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of nonionic polymer of thealkyl aryl polyether alcohol (e.g., tyloxapol (ethoxylatedp-tert-octylphenol formaldehyde polymer)).

The solvent component may comprise 50% phosphatidylcholine in propyleneglycol/ethanol carrier (e.g., Phosal® 50PG), for instance, between about0.01 to about 99.9% of the total weight of the composition; betweenabout 0.1 to about 99%; between about 25 to about 55%; between about 30to about 50%; or between about 35 to about 45%; between about 0.1 toabout 10%; between about 10 to about 20%; between about 20 to about 30%;between about 30 to about 40%; between about 40 to about 45%; betweenabout 40 to about 45%; between about 45 to about 50%; between about 50to about 60%; between about 50 to about 70%; between about 70 to about80%; between about 80 to about 90%; or between about 90 to about 100%.In some variations, 50% phosphatidylcholine in propylene glycol/ethanolcarrier (e.g., Phosal® 50PG) comprises less than about 10% (w/w) of theliquid formulation. In some variations, 50% phosphatidylcholine inpropylene glycol/ethanol carrier (e.g., Phosal® 50PG) comprises lessthan about 5% (w/w) of the liquid formulation. In some variations, 50%phosphatidylcholine in propylene glycol/ethanol carrier (e.g., Phosal®50PG) comprises about 3.12% (w/w) of the liquid formulation.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of 50% phosphatidylcholine in propyleneglycol/ethanol carrier (e.g., Phosal® 50PG). In some variations theliquid formulations described herein contain no greater than about 250μl, no greater than about 200 μl, no greater than about 150 μl, nogreater than about 125 μl, no greater than about 100 μl, no greater thanabout 75 μl, no greater than about 50 μl, no greater than about 25 μl,no greater than about 20 μl, no greater than about 15 μl, no greaterthan about 10 μl, no greater than about 7.5 μl, no greater than about 5μl, no greater than about 2.5 μl, no greater than about 1.0 μl, or nogreater than about 0.5 μl of 50% phosphatidylcholine in propyleneglycol/ethanol carrier (e.g., Phosal® 50PG).

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of 50% phosphatidylcholinein propylene glycol/ethanol carrier (e.g., Phosal® 50PG).

The solvent component may comprise propylene glycol monolaurate, forinstance, between about 0.01 to about 99.9% of the total weight of thecomposition; between about 0.1 to about 99%; between about 25 to about55%; between about 30 to about 50%; or between about 35 to about 45%;between about 0.1 to about 10%; between about 10 to about 20%; betweenabout 20 to about 30%; between about 30 to about 40%; between about 40to about 45%; between about 40 to about 45%; between about 45 to about50%; between about 50 to about 60%; between about 50 to about 70%;between about 70 to about 80%; between about 80 to about 90%; or betweenabout 90 to about 100%. In some variations, propylene glycol monolauratecomprises less than about 10% (w/w) of the liquid formulation. In somevariations, propylene glycol monolaurate comprises less than about 5%(w/w) of the liquid formulation. In some variations, propylene glycolmonolaurate comprises about 3.12% (w/w) of the liquid formulation.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of propylene glycol monolaurate. In somevariations the liquid formulations described herein contain no greaterthan about 250 μl, no greater than about 200 μl, no greater than about150 μl, no greater than about 125 μl, no greater than about 100 μl, nogreater than about 75 μl, no greater than about 50 μl, no greater thanabout 25 μl, no greater than about 20 μl, no greater than about 15 μl,no greater than about 10 μl, no greater than about 7.5 μl, no greaterthan about 5 μl, no greater than about 2.5 μl, no greater than about 1.0μl, or no greater than about 0.5 μl of propylene glycol monolaurate.

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of propylene glycolmonolaurate.

The solvent component may comprise propylene glycoldicaprylate/dicaprate, for instance, between about 0.01 to about 99.9%of the total weight of the composition; between about 0.1 to about 99%;between about 25 to about 55%; between about 30 to about 50%; or betweenabout 35 to about 45%; between about 0.1 to about 10%; between about 10to about 20%; between about 20 to about 30%; between about 30 to about40%; between about 40 to about 45%; between about 40 to about 45%;between about 45 to about 50%; between about 50 to about 60%; betweenabout 50 to about 70%; between about 70 to about 80%; between about 80to about 90%; or between about 90 to about 100%. In some variations,propylene glycol dicaprylate/dicaprate comprises less than about 10%(w/w) of the liquid formulation. In some variations, propylene glycoldicaprylate/dicaprate comprises less than about 5% (w/w) of the liquidformulation. In some variations, propylene glycol dicaprylate/dicapratecomprises about 3.12% (w/w) of the liquid formulation.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of propylene glycol dicaprylate/dicaprate. Insome variations the liquid formulations described herein contain nogreater than about 250 μl, no greater than about 200 μl, no greater thanabout 150 μl, no greater than about 125 μl, no greater than about 100μl, no greater than about 75 μl, no greater than about 50 μl, no greaterthan about 25 μl, no greater than about 20 μl, no greater than about 15μl, no greater than about 10 μl, no greater than about 7.5 μl, nogreater than about 5 μl, no greater than about 2.5 μl, no greater thanabout 1.0 μl, or no greater than about 0.5 μl of propylene glycoldicaprylate/dicaprate.

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of propylene glycoldicaprylate/dicaprate.

The solvent component may comprise macrogol 15 hydroxystearate, forinstance, between about 0.01 to about 99.9% of the total weight of thecomposition; between about 0.1 to about 99%; between about 25 to about55%; between about 30 to about 50%; or between about 35 to about 45%;between about 0.1 to about 10%; between about 10 to about 20%; betweenabout 20 to about 30%; between about 30 to about 40%; between about 40to about 45%; between about 40 to about 45%; between about 45 to about50%; between about 50 to about 60%; between about 50 to about 70%;between about 70 to about 80%; between about 80 to about 90%; or betweenabout 90 to about 100%. In some variations, macrogol 15 hydroxystearatecomprises less than about 10% (w/w) of the liquid formulation. In somevariations, macrogol 15 hydroxystearate comprises less than about 5%(w/w) of the liquid formulation. In some variations, macrogol 15hydroxystearate comprises about 3.12% (w/w) of the liquid formulation.

In some variations the liquid formulations described herein contain nogreater than about 250 μl of macrogol 15 hydroxystearate. In somevariations the liquid formulations described herein contain no greaterthan about 250 μl, no greater than about 200 μl, no greater than about150 μl, no greater than about 125 μl, no greater than about 100 μl, nogreater than about 75 μl, no greater than about 50 μl, no greater thanabout 25 μl, no greater than about 20 μl, no greater than about 15 μl,no greater than about 10 μl, no greater than about 7.5 μl, no greaterthan about 5 μl, no greater than about 2.5 μl, no greater than about 1.0μl, or no greater than about 0.5 μl of macrogol 15 hydroxystearate.

In some variations, the liquid formulations described herein contain nogreater than about 250 mg, no greater than about 200 mg, no greater thanabout 150 mg, no greater than about 125 mg, no greater than about 100mg, no greater than about 75 mg, no greater than about 50 mg, no greaterthan about 25 mg, no greater than about 20 mg, no greater than about 15mg, no greater than about 10 mg, no greater than about 7.5 mg, nogreater than about 5 mg, no greater than about 2.5 mg, no greater thanabout 1.0 mg, or no greater than about 0.5 mg of macrogol 15hydroxystearate.

In some variations, the liquid formulation contains a solvent componentcomprising about 10.5% of NMP, about 73.7% of propylene glycol, andabout 15.8% of water. In some variations, the liquid formulationcontains a solvent component comprising about 38.9% of NMP, about 38.9%of propylene glycol, and about 22.2% of water. In some variations, theliquid formulation contains a solvent component comprising about 10.5%of NMP, about 73.7% of PEG600, and about 15.8% of water. In somevariations, the liquid formulation contains a solvent componentcomprising about 10.5% of DMA, about 73.7% of propylene glycol, andabout 15.8% of water. In some variations, the liquid formulationcontains a solvent component comprising about 10.5% of DMSO, about 73.7%of propylene glycol, and about of 15.8% water. In some variations, theliquid formulation contains a solvent component comprising about 27.9%of NMP, about 39% of PEG400, and about 28.4% of water. In somevariations, the liquid formulation further contains a therapeutic agent.In some variations, the therapeutic agent is less than or equal to about5% of the total weight of the liquid formulation. In some variations,the therapeutic agent is less than any of about 5%, 4%, 3%, 2%, or 1% ofthe total weight of the liquid formulation. In some variations, liquidformulation is administered by subconjunctival, subtenon, and/orintravitreal injection. In some variations, the liquid formulation isused to treat or prevent an ocular disease. In some variations, theocular disease is wet AMD. In some variations, the ocular disease is dryAMD. In some variations, the ocular disease is macular edema.

In some variations, the liquid formulation contains about 8.35% ofpolyethoxylated castor oil (e.g., Cremophor (PEG 35 castor oil)), about8.37% of monoglycerides and/or diglycerides of caprylic acid (e.g.,Capmul MCM (C8)), about 0.07% of therapeutic agent, and about 83.21% ofaqueous liquid (e.g., water or saline). In some variations, the liquidformulation contains about 8.15% of monoglycerides and/or diglyceridesof caprylic acid (e.g., Capmul MCM (C8)), about 8.12% of nonionicpolymer of the alkyl aryl polyether alcohol (e.g., tyloxapol(ethoxylated p-tert-octylphenol formaldehyde polymer)), about 0.09% oftherapeutic agent, about 0.35% of Ethanol, and about 83.29% of aqueousliquid (e.g., water or saline). In some variations, the liquidformulation contains about 6.81% of nonionic polymer of the alkyl arylpolyether alcohol (e.g., tyloxapol (ethoxylated p-tert-octylphenolformaldehyde polymer)), about 6.81% of monoglycerides and/ordiglycerides of caprylic acid (e.g., Capmul MCM (C8)), about 0.08%therapeutic agent, about 3.12% of 50% phosphatidylcholine in propyleneglycol/ethanol carrier (e.g., Phosal® 50PG), and about 83.21% aqueousliquid (e.g., water or saline). In some variations, the liquidformulation contains about 7.18% of polyethoxylated castor oil (e.g.,Cremophor (PEG 35 castor oil)), about 9.47% of monoglycerides and/ordiglycerides of caprylic acid (e.g., Capmul MCM (C8)), about 0.17% oftherapeutic agent, and about 83.18% of aqueous liquid (e.g., water orsaline). In some variations the therapeutic agent is rapamycin. In somevariations, the therapeutic agent is dasatinib. In some variations,liquid formulation is topically administered. In some variations, theliquid formulation is administered as eye drops. In some variations, theliquid formulation is used to treat or prevent an ocular disease. Insome variations, the ocular disease is wet AMD. In some variations, theocular disease is dry AMD. In some variations, the ocular disease ismacular edema.

In some variations, any of the liquid formulation described herein maybe diluted. In some variations, the liquid formulation may be dilutedany of about 5, about 4, about 3, about 2, about 1.9, about 1.8, about1.7, about 1.6, about 1.5, about 1.4, about 1.3, about 1.2, about 1.1fold. In some variations, the liquid formulation is diluted 1.5 fold. Insome variations, the liquid formulation is dilute about 1.4 fold. Insome variations, the liquid formulation is diluted with an aqueousliquid (including, but not limited to, water or saline). In somevariations, the liquid formulation is diluted with saline. In somevariations, the liquid formulation is diluted with water. Fold dilution,as used herein, refers to the final total volume of the diluted liquidformulation divided by the total volume of undiluted liquid formulationadded to make the dilution. For example, a 1.5 fold dilution wouldresult if 500 μl of an undiluted liquid formulation was added to 250 μlof saline to result in a final total volume of the diluted liquidformulation of 750 μl (750 μl of final total volume of the dilutedliquid formulation/500 μl of the total volume of undiluted liquidformulation=1.5). In some variations, the dilution is a solution. Insome variations, the solution is a clear solution. In some variations,the dilution is a suspension. In some variations, the suspension is amilky suspension. In some variations, the dilution is made prior todistribution to the patient or physician. In some variations, thedilution is made by the patient or physician. In some variations, thedilution is made prior to administration. In some variations, the entirediluted liquid formulation is administered to the subject. In somevariations, a portion of the diluted liquid formulation is administeredto the subject. In some variations, a portion of the diluted liquidformulation equal to the volume of the undiluted liquid formulation isadministered to the subject.

Some examples and variations of liquid formulations described hereinwere prepared and are listed in Table 1. Depending on their type, thelisted formulations are denoted one or more of solutions (“S”),suspensions (“SP”), emulsions (“E”) or in situ gelling (“ISG”). Medianparticle size is listed for some of the suspensions. As describedherein, some liquid formulations form a non-dispersed mass after, forexample, injection into an aqueous environment such as the vitreous ofan eye. For those formulations injected into the vitreous of a rabbiteye, the right-hand column of Table 1 indicates whether or not anon-dispersed mass (NDM) formed after a specified volume was injectedinto the vitreous of the rabbit eye.

The following references, each of which is incorporated herein byreference in its entirety, show one or more formulations, including butnot limited to rapamycin formulations, and which describe use ofrapamycin at various doses and other therapeutic agents for treatingvarious diseases or conditions: U.S. 60/651,790, filed Feb. 9, 2005,titled FORMULATIONS FOR OCULAR TREATMENT; U.S. 60/664,040, filed Feb. 9,2005, titled LIQUID FORMULATIONS FOR TREATMENT OF DISEASES ORCONDITIONS; U.S. 60/664,119, filed Mar. 21, 2005, titled DRUG DELIVERYSYSTEMS FOR TREATMENT OF DISEASES OR CONDITIONS; U.S. 60/664,306, filedMar. 21, 2005, titled IN SITU GELLING FORMULATIONS AND LIQUIDFORMULATIONS FOR TREATMENT OF DISEASES OR CONDITIONS; U.S. Ser. No.11/351,844, filed Feb. 9, 2006, titled FORMULATIONS FOR OCULARTREATMENT; U.S. Ser. No. 11/351,761, filed Feb. 9, 2006, titled LIQUIDFORMULATIONS FOR TREATMENT OF DISEASES OR CONDITIONS; US 2005/0187241,and US 2005/0064010.

Liquid Formulations which Form a Non-Dispersed Mass

One class of liquid formulations described herein forms a non-dispersedmass when placed in an aqueous medium. As used herein, a “non-dispersedmass” refers to the structure formed or shape assumed when the liquidformulation is placed into an environment, relative to the environmentin which it is placed. Generally, a non-dispersed mass of a liquidformulation is anything other than a homogeneous distribution of theliquid formulation in the surrounding medium. The non-dispersed massmay, for instance, be indicated by visually inspecting the administeredliquid formulation and characterizing its appearance relative to thesurrounding medium.

In some variations, the aqueous medium is water. In some variations, thewater is deionized, distilled, sterile, or tap water, including but notlimited to tap water available at the place of business of MacuSight inUnion City, Calif.

In some variations, the aqueous medium is an aqueous medium of asubject. In some variations the aqueous medium is an aqueous medium ofthe eye of a subject, including but not limited to the vitreous of aneye of a subject. In some variations the subject is a human subject. Insome variations the subject is a rabbit.

In some variations the liquid formulation forms a non-dispersed masswhen exposed to a certain temperature or range of temperatures,including but not limited to about room temperature, about ambienttemperature, about 30° C., about 37° C., or about the temperature of theaqueous medium of the subject.

In some variations the liquid formulation forms a non-dispersed masswhen exposed to a certain pH or range of pH, including but not limitedto a pH between about 6 and about 8.

In some variations, the non-dispersed mass comprises a gel or gel-likesubstance.

In some variations, the non-dispersed mass comprises a polymer matrix.In some variations, the non-dispersed mass comprises a polymer matrix inwhich a therapeutic agent is dispersed.

The liquid formulations described herein may generally be of anygeometry or shape after administration to a subject or the eye of asubject, including but not limited to a human subject. In somevariations, the non-dispersed mass is between about 0.1 and about 5 mm.In some variations, the non-dispersed mass is between about 1 and about3 mm. The non-dispersed mass-forming liquid formulations may, forinstance, appear as a compact spherical mass when administered to thevitreous. In some instances, the liquid formulation may appear as anon-dispersed mass relative to the surrounding medium, wherein thenon-dispersed mass is less clearly defined and the geometry is moreamorphous than spherical.

The non-dispersed mass-forming liquid formulations described herein mayform a non-dispersed mass immediately upon placement in the medium orthe non-dispersed mass may form some period of time after placement ofthe liquid formulation. In some variations the non-dispersed mass formsover the course of about 1, about 2, about 3, about 4, about 5, about 6,or about 7 days. In some variations the non-dispersed mass forms overthe course of about 1 week, about 2 weeks, or about 3 weeks.

In some variations, the liquid formulations described herein that form anon-dispersed mass appear as a milky or whitish colored semi-contiguousor semi-solid non-dispersed mass relative to the medium in which it isplaced.

One liquid formulation described herein forms a non-dispersed mass whichhas the form of a solid depot when the formulation is injected into anyor all of water, the vitreous of a rabbit eye, or between the sclera andthe conjunctiva of a rabbit eye. One liquid formulation described hereinforms a non-dispersed mass which has the form of a semi-solid when theformulation is injected into any or all of water, the vitreous of arabbit eye, or between the sclera and the conjunctiva of a rabbit eye.One liquid formulation described herein forms a non-dispersed mass whichhas the form of a polymeric matrix when the formulation is injected intoany or all of water, the vitreous of a rabbit eye, or between the scleraand the conjunctiva of a rabbit eye. One liquid formulation describedherein forms a non-dispersed mass which has the form of a gel, ahydrogel, or a gel-like substance when the formulation is injected intoany or all of water, the vitreous of a rabbit eye, or between the scleraand the conjunctiva of a rabbit eye.

In some variations described herein the liquid formulation forms anon-dispersed mass relative to a surrounding medium where thesurrounding medium is aqueous. An “aqueous medium” or “aqueousenvironment” is one that contains at least about 50% water. Examples ofaqueous media include but are not limited to water, the vitreous,extracellular fluid, conjunctiva, sclera, between the sclera and theconjunctiva, aqueous humor, gastric fluid, and any tissue or body fluidcomprised of at least about 50% of water. Aqueous media include but arenot limited to gel structures, including but not limited to those of theconjunctiva and sclera.

In some variations, the liquid formulations described herein form anon-dispersed mass when a test volume of the liquid formulation isplaced in the vitreous of a rabbit eye. In some variations the testvolume administered to a rabbit eye, and the test volume is equal to thevolume of the liquid formulation administered to a subject's, includingbut not limited to a human subject's eye.

In some variations, the test volume administered to a rabbit eye isequal to the volume administered to the subject's eye multiplied by ascale factor, and the scale factor is equal to the average volume of arabbit eye divided by the average volume of a subject eye. The “averagevolume” of an eye, as used herein, refers to the average volume of aneye of a member of similar age of the species under considerationgenerally, as opposed to the average volume of any particularindividual's eye.

In some variations, the test volume administered to the rabbit eye isbetween about 10 μl and about 50 μl. In some variations, the test volumeadministered to the rabbit eye is between about 1 μl and about 30 μl. Insome variations, the test volume administered to the rabbit eye isbetween about 50 μl and about 100 μl. In some variations, the testvolume administered to the rabbit eye is between about 25 μl and about75 μl. In some variations, the test volume administered to the rabbiteye is about 30 μl.

In some variations, the liquid formulation that forms a non-dispersedmass when placed in the medium may comprises a therapeutic agent oragents with a concentration of between about 0.01% and about 10% byweight of the total, and a solvent between about 10% and about 99% byweight of the total. In some variations the formulation furthercomprises a solubilizing agent including but not limited to asurfactant. In some variations the liquid formulation further comprisesa stabilizing agent, excipient, adjuvant, or antioxidant, etc., betweenabout 0 and about 40% by weight of the total. In some variations, thetherapeutic agent is about 5% by weight of the total, and the solventcomponent is about 95% by weight of the total.

Whether a liquid formulation exhibits a non-dispersed mass relative to asurrounding medium when present in a subject, including but not limitedto a human subject or the eye of a subject may be determined by, forinstance, mixing a therapeutic agent with a solvent, administering it tothe vitreous of an eye of a subject, including but not limited to ahuman subject, and comparing the liquid formulation to the surroundingmedium.

One liquid formulation that may be used for treating, preventing,inhibiting, delaying the onset of, or causing the regression of thediseases and conditions of a subject, including but not limited to ahuman subject, is a liquid formulation that forms a non-dispersed masswhen placed into the vitreous of a rabbit eye. When used for treating,preventing, inhibiting, delaying the onset of, or causing the regressionof the disease or condition of the subject, the liquid formulation isadministered to the subject. The liquid formulation may or may not forma non-dispersed mass in the subject. One liquid formulation describedherein forms a non-dispersed mass when administered to a subject andforms a non-dispersed mass when administered to a rabbit eye.

Without being bound by theory, it is believed that the low solubility ofrapamycin in the vitreous contributes to the formation of anon-dispersed mass by some rapamycin-containing liquid formulationsdescribed herein. The vitreous is a clear gel composed almost entirelyof water (up to 99%). Without being bound by theory, it is believed thatas rapamycin in an injected formulation contacts the vitreous, therapamycin precipitates.

Without being bound by theory, factors believed to affect the formationof and geometry of a non-dispersed mass include the concentration ofrapamycin in the formulation, the viscosity of the formulation, ethanolcontent of the formulation, and the volume of injection. It is believedthat maintaining a higher local concentration of rapamycin afterinjection of the formulation favors formation of a non-dispersed mass,as opposed to a lower local concentration of rapamycin after injectionof the formulation. As volume is increased for a given dose, formationof a non-dispersed mass may become less favorable. Formation of anon-dispersed mass may become more favorable as rapamycin concentrationis increased and/or as viscosity is increased. Ethanol content affectsboth the solubility of the rapamycin in the formulation and theviscosity of the formulation.

In one comparison, 100 μl of a solution of 0.4% rapamycin, 4.0% ethanol,and 95.6% PEG 400 (a 400 μg dose) did not form a non-dispersed massafter injection into a rabbit eye. In contrast, 20 μl of a solution of2.00% rapamycin, 4.0% ethanol, and 94% PEG 400 (also a 400 μg dose)formed a compact spherical non-dispersed mass after injection into arabbit eye.

Without being bound by theory, in the latter example, it is hypothesizedthat formation of the non-dispersed mass occurred as depicted in FIGS.1A-1C and described as follows. Upon injection, due to its viscosity theliquid formulation formed a spherical globule 100 within the vitreous110. Ethanol then diffused out of this globule, resulting in localizedprecipitation 120 of the rapamycin within the globule. Eventually, thepolyethylene glycol also diffused out of the globule to leave a solid,compact non-dispersed mass of rapamycin 130.

In some variations, the non-dispersed masses described herein consistsof at least about 20%, at least about 30%, at least about 40%, at leastabout 50%, at least about 60%, at least about 70%, at least about 80%,at least about 90%, or at least about 95% by volume of therapeutic agentwhen injected into the vitreous of a rabbit eye.

In some variations, upon formation a non-dispersed mass comprisingrapamycin, for example, delivers the drug continuously at approximatelya constant rate for an extended period of time. Without being bound bytheory, it is believed that delivery of rapamycin from a non-dispersedmass in the vitreous depends on dissolution of the rapamycin in thevitreous, which depends in turn on clearance of the drug from thevitreous to other tissues. Without being bound by theory, this releaseprocess is believed to maintain a steady-state concentration ofrapamycin in the vitreous.

In some variations, formation of a non-dispersed mass reduces thetoxicity of the injected liquid formulation compared to an equivalentdose that did not form a non-dispersed mass. In variations in which aliquid formulation injected into the vitreous does not form anon-dispersed mass, the drug (e.g., rapamycin) appears to disperse inthe vitreous body. In some variations this may interfere with vision.

In some variations, liquid formulations that are suspensions form anon-dispersed mass upon injection into the vitreous. Formation of anon-dispersed mass from an injected suspension may become more favorableas the suspension particle size increases.

In some variations, it is believed that the liquid formulations willform a visually observable non-dispersed mass when injected into the eyeof a subject, including but not limited to a human subject.

In some variations, liquid formulations are believed to formnon-dispersed masses when injected subconjunctivally. In some variationsit is believed that when subconjunctivally administered the liquidformulation forms a depot in the scleral tissue. That is, it is believedthat the therapeutic agent is absorbed into the sclera proximate to theinjection site and forms a local concentration of drug in the sclera.

In Situ Gelling Formulations

Described herein are non-dispersed mass-forming liquid formulationswhich form a gel or gel-like substance when placed in an aqueous medium.In some variations, the non-dispersed mass comprises a gel; in somevariations the gel is a hydrogel.

An “in situ gelling formulation,” as used herein, refers to a liquidformulation which forms a gel-like non-dispersed mass when the liquidformulation is placed in an aqueous medium, including but not limited toaqueous media that are water, the vitreous of a rabbit eye, and betweenthe sclera and the conjunctiva of a rabbit eye. In some variations, anin situ gelling formulation forms a gel-like non-dispersed mass whenplaced in tap water.

In some variations, the in situ gelling formulation is a suspensionprior to placement in an aqueous medium, and forms a gel in situ uponplacement in an aqueous medium. In some variations, the in situ gellingformulation is a solution prior to placement in an aqueous medium, andforms a gel in situ upon placement in an aqueous medium. In somevariations, the in situ gelling formulation is an emulsion prior toplacement in an aqueous medium, and forms a gel in situ upon placementin an aqueous medium. In some variations a gel-like non-dispersed massforms after placement of the in situ gelling formulation into an aqueousmedium, including but not limited to any or all of water, the vitreous,or between the sclera and the conjunctiva of an eye. In some variations,the in situ gel is formed of a polymer matrix. In some variations atherapeutic agent is dispersed in the polymer matrix.

Described herein are in situ gelling formulations which may be used fortreating, preventing, inhibiting, delaying the onset of, or causing theregression of the diseases and conditions of a subject including but notlimited to a human subject. When used for treating, preventing,inhibiting, delaying the onset of, or causing the regression of thedisease or condition of the subject, the in situ gelling formulation isadministered to the subject. One liquid formulation described hereincomprises an in situ gelling formulation which forms a non-dispersedmass when administered to a subject and forms a non-dispersed mass whenadministered to a rabbit eye.

In some variations, the in situ gelling formulation comprises one ormore polymers. Described herein are various types of polymers, includingpolymers which are solvents, polymers which are solubilizing agents,polymers which are release modifying agents, polymers which arestabilizing agents, etc. In some variations, any combination of polymersis used wherein the polymers when combined with the therapeutic agentform any or all of a non-dispersed mass, a gel, a hydrogel, or polymericmatrix when placed in an aqueous medium, including but not limited toany or all of water, the vitreous, or between the sclera and theconjunctiva.

In some variations, the in situ gelling formulation delivers extendedrelease of therapeutic agents to a subject when administered to thesubject.

In some variations, the liquid formulation comprises a therapeutic agentand a plurality of polymers, wherein one of the polymers is apolymethacrylate. Polymethacrylates are known by various names and areavailable in various preparations, including but not limited topolymeric methacrylates, methacrylic acid-ethyl acrylate copolymer(1:1), methacrylic acid-ethyl acrylate copolymer (1:1) dispersion 30percent, methacrylic acid-methyl methacrylate copolymer (1:1),methacrylic acid-methyl methacrylate copolymer (1:2), acidummethacrylicum et ethylis acrylas polymerisatum 1:1, acidum methacrylicumet ethylis acrylas polymerisatum 1:1 dispersio 30 per centum, acidummethacrylicum et methylis methacrylas polymerisatum 1:1, acidummethacrylicum et methylis methacrylas polymerisatum 1:2, USPNF: ammoniomethacrylate copolymer, methacrylic acid copolymer, methacrylic acidcopolymer dispersion.

In some variations, one of the polymers is polyvinylpyrrolidone.Polyvinylpyrrolidone is known by various names and is available invarious preparations, including but not limited to povidone, povidonum,kollidon; plasdone; poly[1-(2-oxo-1-pyrrolidinyl)ethylene]; polyvidone;PVP; 1-vinyl-2-pyrrolidinone polymer, and 1-Ethenyl-2-pyrrolidinonehomopolymer.

One liquid formulation described herein comprises a therapeutic agentand a solvent component. The solvent component may comprise a singlesolvent or a combination of solvents.

In some variations, the solvent is glycerin, dimethylsulfoxide,N-methylpyrrolidone, ethanol, isopropyl alcohol, polyethylene glycol ofvarious molecular weights, including but not limited to PEG 300 and PEG400, or propylene glycol, or a mixture of one or more thereof.

In some variations, the solvent is polyethylene glycol. Polyethyleneglycol is known by various names and is available in variouspreparations, including but not limited to macrogels, macrogel 400,macrogel 1500, macrogel 4000, macrogel 6000, macrogel 20000, macrogola,breox PEG; carbowax; carbowax sentry; Hodag PEG; Lipo; Lipoxol; LutrolE; PEG; Pluriol E; polyoxyethylene glycol, andα-Hydro-ω-hydroxy-poly(oxy-1,2-ethanediyl).

Compositions and Liquid Formulations for Delivery of Therapeutic Agents

The compositions and liquid formulations described herein may be used todeliver amounts of the therapeutic agents effective for treating,preventing, inhibiting, delaying on set of, or causing the regression ofthe diseases and conditions described in the Diseases and Conditionssection. In some variations the compositions and liquid formulationsdescribed herein deliver one or more therapeutic agents over an extendedperiod of time.

An “effective amount,” which is also referred to herein as a“therapeutically effective amount,” of a therapeutic agent foradministration as described herein is that amount of the therapeuticagent that provides the therapeutic effect sought when administered tothe subject, including but not limited to a human subject. The achievingof different therapeutic effects may require different effective amountsof therapeutic agent. For example, the therapeutically effective amountof a therapeutic agent used for preventing a disease or condition may bedifferent from the therapeutically effective amount used for treating,inhibiting, delaying the onset of, or causing the regression of thedisease or condition. In addition, the therapeutically effective amountmay depend on the age, weight, and other health conditions of thesubject as is well know to those versed in the disease or conditionbeing addressed. Thus, the therapeutically effective amount may not bethe same in every subject to which the therapeutic agent isadministered.

An effective amount of a therapeutic agent for treating, preventing,inhibiting, delaying the onset of, or causing the regression of aspecific disease or condition is also referred to herein as the amountof therapeutic agent effective to treat, prevent, inhibit, delay theonset of, or cause the regression of the disease or condition.

To determine whether a level of therapeutic agent is a “therapeuticallyeffective amount” to treat, prevent, inhibit, delay on set of, or causethe regression of the diseases and conditions described in the Diseasesand Conditions section, liquid formulations may be administered inanimal models for the diseases or conditions of interest, and theeffects may be observed. In addition, dose ranging human clinical trialsmay be conducted to determine the therapeutically effective amount of atherapeutic agent.

Generally, the therapeutic agent may be formulated in any composition orliquid formulation capable of delivery of a therapeutically effectiveamount of the therapeutic agent to a subject or to the eye of a subjectfor the required delivery period. Compositions include liquidformulations.

Solubilization of Therapeutic Agents

One composition or liquid formulation that may be used is a compositionor liquid formulation in which the therapeutic agent is dissolved in asolvent component. Generally, any solvent which has the desired effectmay be used in which the therapeutic agent dissolves. In some variationsthe solvent is aqueous. In some variations the solvent is non-aqueous.An “aqueous solvent” is a solvent that contains at least about 50%water.

Generally, any concentration of solubilized therapeutic agent that hasthe desired effect can be used. The solvent component may be a singlesolvent or may be a mixture of solvents. The solvent component may be asingle solvent or may be a mixture of solvents. Solvents and types ofsolutions are well known to those versed in such drug deliverytechnologies. See for example, Remington: The Science and Practice ofPharmacy, Twentieth Edition, Lippincott Williams & Wilkins; 20th edition(Dec. 15, 2000); Ansel's Pharmaceutical Dosage Forms and Drug DeliverySystems, Eighth Edition, Lippincott Williams & Wilkins (August 2004);Handbook Of Pharmaceutical Excipients 2003, American PharmaceuticalAssociation, Washington, D.C., USA and Pharmaceutical Press, London, UK;and Strickley, solubilizing Excipients in Oral and InjectableFormulations, Pharmaceutical Research, Vol. 21, No. 2, February 2004.

As noted previously, some solvents may also serve as solubilizingagents.

Solvents that may be used include but are not limited to DMSO, ethanol,methanol, isopropyl alcohol; castor oil, propylene glycol, glycerin,polysorbate 80, benzyl alcohol, dimethyl acetamide (DMA), dimethylformamide (DMF), triacetin, diacetin, corn oil, acetyl triethyl citrate(ATC), ethyl lactate, glycerol formal, ethoxy diglycol (Transcutol,Gattefosse), tryethylene glycol dimethyl ether (Triglyme), dimethylisosorbide (DMI), γ-butyrolactone, N-Methyl-2-pyrrolidinone (NMP),polyethylene glycol of various molecular weights, including but notlimited to PEG 300 and PEG 400, and polyglycolated capryl glyceride(Labrasol, Gattefosse), combinations of any one or more of theforegoing, or analogs or derivatives of any one or more of theforegoing.

In some variations, the solvent is a polyethylene glycol. Polyethyleneglycol is known by various names and is available in variouspreparations, including but not limited to macrogels, macrogel 400,macrogel 1500, macrogel 4000, macrogel 6000, macrogel 20000, macrogola,breox PEG; carbowax; carbowax sentry; Hodag PEG; Lipo; Lipoxol; LutrolE; PEG; Pluriol E; polyoxyethylene glycol, andα-Hydro-ω-hydroxy-poly(oxy-1,2-ethanediyl).

In some variations the polyethylene glycol is a liquid PEG, and is oneor more of PEG 300 or PEG 400.

Other solvents include an amount of a C₆-C₂₄ fatty acid sufficient tosolubilize a therapeutic agent.

Phospholipid solvents may also be used, such as lecithin,phosphatidylcholine, or a mixture of various diglycerides of stearic,palmitic, and oleic acids, linked to the choline ester of phosphoricacid; hydrogenated soy phosphatidylcholine (HSPC),distearoylphosphatidylglycerol (DSPG),L-α-dimyristoylphosphatidylcholine (DMPC),L-α-dimyristoylphosphatidylglycerol (DMPG).

Further examples of solvents include, for example, components such asalcohols, propylene glycol, polyethylene glycol of various molecularweights, propylene glycol esters, propylene glycol esterified with fattyacids such as oleic, stearic, palmic, capric, linoleic, etc; mediumchain mono-, di-, or triglycerides, long chain fatty acids, naturallyoccurring oils, and a mixture thereof. The oily components for thesolvent system include commercially available oils as well as naturallyoccurring oils. The oils may further be vegetable oils or mineral oils.The oils can be characterized as non-surface active oils, whichtypically have no hydrophile lipophile balance value. Commerciallyavailable substances comprising medium chain triglycerides include, butare not limited to, Captex 100, Captex 300, Captex 355, Miglyol 810,Miglyol 812, Miglyol 818, Miglyol 829, and Dynacerin 660. Propyleneglycol ester compositions that are commercially available encompassCaptex 200 and Miglyol 840, and the like. The commercial product, CapmulMCM, comprises one of many possible medium chain mixtures comprisingmonoglycerides and diglycerides.

Other solvents include naturally occurring oils such as peppermint oil,and seed oils. Exemplary natural oils include oleic acid, castor oil,safflower seed oil, soybean oil, olive oil, sunflower seed oil, sesameoil, and peanut oil. Soy fatty acids may also be used. Examples of fullysaturated non-aqueous solvents include, but are not limited to, estersof medium to long chain fatty acids (such as fatty acid triglycerideswith a chain length of about C₆ to about C₂₄). Hydrogenated soybean oiland other vegetable oils may also be used. Mixtures of fatty acids maybe split from the natural oil (for example coconut oil, palm kernel oil,babassu oil, or the like) and refined. In some embodiments, medium chain(about C₈ to about C₁₂) triglycerides, such as caprilyic/caprictriglycerides derived from coconut oil or palm seed oil, may be used.Medium chain mono- and diglycerides may also be used. Other fullysaturated non-aqueous solvents include, but are not limited to,saturated coconut oil (which typically includes a mixture of lauric,myristic, palmitic, capric and caproic acids), including those soldunder the Miglyol™ trademark from Huls and bearing trade designations810, 812, 829 and 840). Also noted are the NeoBee™ products sold by DrewChemicals. Non-aqueous solvents include isopropyl myristate. Examples ofsynthetic oils include triglycerides and propylene glycol diesters ofsaturated or unsaturated fatty acids having 6 to 24 carbon atoms suchas, for example hexanoic acid, octanoic (caprylic), nonanoic(pelargonic), decanoic (capric), undecanoic, lauric, tridecanoic,tetradecanoic (myristic), pentadecanoic, hexadecanoic (palmitic),heptadecanoic, octadecanoic (stearic), nonadecanoic, heptadecanoic,eicosanoic, heneicosanoic, docosanoic and lignoceric acids, and thelike. Examples of unsaturated carboxylic acids include oleic, linoleicand linolenic acids, and the like. The non-aqueous solvent can comprisethe mono-, di- and triglyceryl esters of fatty acids or mixed glyceridesand/or propylene glycol mono- or diesters wherein at least one moleculeof glycerol has been esterified with fatty acids of varying carbon atomlength. A non-limiting example of a “non-oil” useful as a solvent ispolyethylene glycol.

Exemplary vegetable oils include cottonseed oil, corn oil, sesame oil,soybean oil, olive oil, fractionated coconut oil, peanut oil, sunfloweroil, safflower oil, almond oil, avocado oil, palm oil, palm kernel oil,babassu oil, beechnut oil, linseed oil, rape oil and the like. Mono-,di-, and triglycerides of vegetable oils, including but not limited tocorn, may also be used.

Polyvinyl pyrrolidone (PVP), cross-linked or not, may also be used as asolvent. Further solvents include but are not limited to C₆-C₂₄ fattyacids, oleic acid, Imwitor 742, Capmul, F68, F68 (Lutrol), PLURONICSincluding but not limited to PLURONICS F108, F127, and F68, Poloxamers,Jeffamines), Tetronics, F127; cyclodextrins such as α-cyclodextrin,β-cyclodextrin, hydroxypropyl-β-cyclodextrin,sulfobutylether-β-cyclodextrin (Captisol); CMC, polysorbitan 20,Cavitron, polyethylene glycol of various molecular weights including butnot limited to PEG 300 and PEG 400.

Beeswax and d-α-tocopherol (Vitamin E) may also be used as solvents.

In some variations, the solvent is N-methylpyrrolidone (NMP),dimethyl-acetamine (DMA), dimethyl sulfoxide (DMSO), propylene glycol(PG), polyethylene glycol 600 (PEG 600), polyethylene glycol 400 (PEG400), ethanol, or a mixture of one or more thereof. In some variations,the solvent comprises a combination of solvents including N-methylpyrrolidone (NMP), dimethyl-acetamine (DMA), or dimethyl sulfoxide(DMSO). In some variations, the solvent comprises a combination ofsolvents including propylene glycol (PG), polyethylene glycol 600 (PEG600), or polyethylene glycol 400 (PEG 400). In some variation, thesolvent may comprise a combination of at least two solvents. In somevariations, the at least two solvents comprising a first solvent such asN-methylpyrrolidone (NMP), dimethyl-acetamine (DMA), or dimethylsulfoxide (DMSO) and a second solvent such as propylene glycol (PG),polyethylene glycol 600 (PEG 600), or polyethylene glycol 400 (PEG 400).In some variations, the solvent may comprise N-methylpyrrolidone (NMP)and propylene glycol (PG). In some variations, the solvent may compriseN-methylpyrrolidone (NMP) and polyethylene glycol 600 (PEG 600). In somevariations, the solvent may comprise dimethyl-acetamine (DMA) andpropylene glycol (PG). In some variations, the solvent component maycomprise dimethyl-acetamine (DMA) and polyethylene glycol 400 (PEG 400).In some variations, the solvent may comprise dimethyl sulfoxide (DMSO)and propylene glycol (PG). In some variations, the solvent may compriseN-methylpyrrolidone (NMP) and polyethylene glycol 400 (PEG 400). In somevariations, the solvent may further comprise ethanol. In somevariations, the solvent may further comprise water. In some variations,the solvent may have a hygroscopicity that is about equal to, less than,or greater than that of polyethylene glycol 400 (PEG 400).

In some variations, the solvent is polyethoxylated castor oil (e.g.,Cremophor (PEG 35 castor oil)), monoglycerides and/or diglycerides ofcaprylic acid (e.g., Capmul MCM (C8)), nonionic polymer of the alkylaryl polyether alcohol (e.g., tyloxapol (ethoxylated p-tert-octylphenolformaldehyde polymer)), Phosal® 50PG, ethanol, or a mixture of one ormore thereof. In some variation, the solvent may comprise a combinationof at least two solvents. In some variations, the at least two solventscomprising a first solvent such as polyethoxylated castor oil (e.g.,Cremophor (PEG 35 castor oil)) or nonionic polymer of the alkyl arylpolyether alcohol (e.g., tyloxapol (ethoxylated p-tert-octylphenolformaldehyde polymer)) and a second solvent such as monoglyceridesand/or diglycerides of caprylic acid (e.g., Capmul MCM (C8)). In somevariations, the solvent may further comprise 50% phosphatidylcholine inpropylene glycol/ethanol carrier (e.g., Phosal® 50PG).

Solvents for use in the liquid formulations can be determined by avariety of methods known in the art, including but not limited to (1)theoretically estimating their solubility parameter values and choosingthe ones that match with the therapeutic agent, using standard equationsin the field; and (2) experimentally determining the saturationsolubility of therapeutic agent in the solvents, and choosing the onesthat exhibit the desired solubility.

Solubilization of Rapamycin

Where the therapeutic agent is rapamycin, solvents that may be used formaking solutions or suspensions of rapamycin include but are not limitedto any solvent described herein, including but not limited to any one ormore of DMSO, glycerin, ethanol, methanol, isopropyl alcohol; castoroil, propylene glycol, polyvinylpropylene, glycerin, polysorbate 80,benzyl alcohol, dimethyl acetamide (DMA), dimethyl formamide (DMF),glycerol formal, ethoxy diglycol (Transcutol, Gattefosse), tryethyleneglycol dimethyl ether (Triglyme), dimethyl isosorbide (DMI),γ-butyrolactone, N-Methyl-2-pyrrolidinone (NMP), polyethylene glycol ofvarious molecular weights, including but not limited to PEG 300 and PEG400, and polyglycolated capryl glyceride (Labrasol, Gattefosse).

Further solvents include but are not limited to C₆-C₂₄ fatty acids,oleic acid, Imwitor 742, Capmul, F68, F68 (Lutrol), PLURONICS includingbut not limited to PLURONICS F108, F127, and F68, Poloxamers,Jeffamines), Tetronics, F127, beta-cyclodextrin, CMC, polysorbitan 20,Cavitron, softigen 767, captisol, and sesame oil.

Other methods that may be used to dissolve rapamycin are described inSolubilization of Rapamycin, P. Simamora et al. Int'l J. Pharma 213(2001) 25-29, the contents of which is incorporated herein in itsentirety.

As a nonlimiting example, rapamycin can be dissolved in 5% DMSO ormethanol in a balanced salt solution. The rapamycin solution can beunsaturated, a saturated or a supersaturated solution of rapamycin. Therapamycin solution can be in contact with solid rapamycin. In onenonlimiting example, rapamycin can be dissolved in a concentration of upto about 400 mg/ml. Rapamycin can also, for example, be dissolved inpropylene glycol esterified with fatty acids such as oleic, stearic,palmic, capric, linoleic, etc.

In some variations, the solvent is N-methylpyrrolidone (NMP),dimethyl-acetamine (DMA), dimethyl sulfoxide (DMSO), propylene glycol(PG), polyethylene glycol 600 (PEG 600), polyethylene glycol 400 (PEG400), ethanol, or a mixture of one or more thereof. In some variations,the solvent comprises a combination of solvents including N-methylpyrrolidone (NMP), dimethyl-acetamine (DMA), or dimethyl sulfoxide(DMSO). In some variations, the solvent comprises a combination ofsolvents including propylene glycol (PG), polyethylene glycol 600 (PEG600), or polyethylene glycol 400 (PEG 400). In some variation, thesolvent may comprise a combination of at least two solvents. In somevariations, the at least two solvents comprising a first solvent such asN-methylpyrrolidone (NMP), dimethyl-acetamine (DMA), or dimethylsulfoxide (DMSO) and a second solvent such as propylene glycol (PG),polyethylene glycol 600 (PEG 600), or polyethylene glycol 400 (PEG 400).In some variations, the solvent may comprise N-methylpyrrolidone (NMP)and propylene glycol (PG). In some variations, the solvent may compriseN-methylpyrrolidone (NMP) and polyethylene glycol 600 (PEG 600). In somevariations, the solvent may comprise dimethyl-acetamine (DMA) andpropylene glycol (PG). In some variations, the solvent component maycomprise dimethyl-acetamine (DMA) and polyethylene glycol 400 (PEG 400).In some variations, the solvent may comprise dimethyl sulfoxide (DMSO)and propylene glycol (PG). In some variations, the solvent may compriseN-methylpyrrolidone (NMP) and polyethylene glycol 400 (PEG 400). In somevariations, the solvent may further comprise ethanol. In somevariations, the solvent may further comprise water. In some variations,the solvent may have a hygroscopicity that is about equal to, less than,or greater than that of polyethylene glycol 400 (PEG 400).

In some variations, the solvent is polyethoxylated castor oil (e.g.,Cremophor (PEG 35 castor oil)), monoglycerides and/or diglycerides ofcaprylic acid (e.g., Capmul MCM (C8)), nonionic polymer of the alkylaryl polyether alcohol (e.g., tyloxapol (ethoxylated p-tert-octylphenolformaldehyde polymer)), Phosal® 50PG, ethanol, or a mixture of one ormore thereof. In some variation, the solvent may comprise a combinationof at least two solvents. In some variations, the at least two solventscomprising a first solvent such as polyethoxylated castor oil (e.g.,Cremophor (PEG 35 castor oil)) or nonionic polymer of the alkyl arylpolyether alcohol (e.g., tyloxapol (ethoxylated p-tert-octylphenolformaldehyde polymer)) and a second solvent such as monoglyceridesand/or diglycerides of caprylic acid (e.g., Capmul MCM (C8)). In somevariations, the solvent may further comprise 50% phosphatidylcholine inpropylene glycol/ethanol carrier (e.g., Phosal® 50PG). In somevariations, the solvent may further comprise ethanol. In somevariations, the solvent may further comprise water.

Many other solvents are possible. Those of ordinary skill in the artwill find it routine to identify solvents for rapamycin given theteachings herein.

Solubilizing Agents

Generally, any solubilizing agent or combination of solubilizing agentsmay be used in the liquid formulations described herein.

In some variations, the solubilizing agent is a surfactant orcombination of surfactants. Many surfactants are possible. Combinationsof surfactants, including combinations of various types of surfactants,may also be used. For instance, surfactants which are nonionic, anionic(i.e. soaps, sulfonates), cationic (i.e. CTAB), zwitterionic, polymericor amphoteric may be used.

Surfactants that can be used may be determined by mixing a therapeuticagent of interest with a putative solvent and a putative surfactant, andobserving the characteristics of the formulation after exposure to amedium.

Examples of surfactants include but are not limited to fatty acid estersor amides or ether analogues, or hydrophilic derivatives thereof;monoesters or diesters, or hydrophilic derivatives thereof; or mixturesthereof; monoglycerides or diglycerides, or hydrophilic derivativesthereof; or mixtures thereof; mixtures having enriched mono- or/anddiglycerides, or hydrophilic derivatives thereof; surfactants with apartially derivatized with a hydrophilic moiety; monoesters or diestersor multiple-esters of other alcohols, polyols, saccharides oroligosaccharides or polysaccharides, oxyalkylene oligomers or polymersor block polymers, or hydrophilic derivatives thereof, or the amideanalogues thereof; fatty acid derivatives of amines, polyamines,polyimines, aminoalcohols, aminosugars, hydroxyalkylamines,hydroxypolyimines, peptides, polypeptides, or the ether analoguesthereof.

Hydrophilic Lipophilic Balance (“HLB”) is an expression of the relativesimultaneous attraction of a surfactant for water and oil (or for thetwo phases of the emulsion system being considered).

Surfactants are characterized according to the balance between thehydrophilic and lipophilic portions of their molecules. Thehydrophilic-lipophilic balance (HLB) number indicates the polarity ofthe molecule in an arbitrary range of 1-40, with the most commonly usedemulsifiers having a value between 1-20. The HLB increases withincreasing hydrophilicity.

Surfactants that may be used include but are not limited to those withan HLB greater than 10, 11, 12, 13 or 14. Examples of surfactantsinclude polyoxyethylene products of hydrogenated vegetable oils,polyethoxylated castor oils or polyethoxylated hydrogenated castor oil,polyoxyethylene-sorbitan-fatty acid esters, polyoxyethylene castor oilderivatives and the like, for example, Nikkol HCO-50, Nikkol HCO-35,Nikkol HCO-40, Nikkol HCO-60 (from Nikko Chemicals Co. Ltd.); Cremophor(from BASF) such as Cremophor RH40, Cremophor RH60, Cremophor EL, TWEENs(from ICI Chemicals) e.g., TWEEN 20, TWEEN 21, TWEEN 40, TWEEN 60, TWEEN80, TWEEN 81, Cremophor RH 410, Cremophor RH 455 and the like.

The surfactant component may be selected from compounds having at leastone ether formed from at least about 1 to 100 ethylene oxide units andat least one fatty alcohol chain having from at least about 12 to 22carbon atoms; compounds having at least one ester formed from at leastabout 1 to 100 ethylene oxide units and at least one fatty acid chainhaving from at least about 12 to 22 carbon atoms; compounds having atleast one ether, ester or amide formed from at least about 1 to 100ethylene oxide units and at least one vitamin or vitamin derivative; andcombinations thereof consisting of no more than two surfactants.

Other examples of surfactants include Lumulse GRH-40, TGPS,Polysorbate-80 (TWEEN-80), Polysorbate-20 (TWEEN-20), polyoxyethylene(20) sorbitan mono-oleate), glyceryl glycol esters, polyethylene glycolesters, polyglycolyzed glycerides, and the like, or mixtures thereof;polyethylene sorbitan fatty acid esters, polyoxyethylene glycerolesters, such as Tagat TO, Tagat L, Tagat I, tagat I2 and Tagat 0(commercially available from Goldschmidt Chemical Co., Essen, Germany);ethylene glycol esters, such as glycol stearate and distearate;propylene glycol esters, such as propylene glycol myristate; glycerylesters of fatty acids, such as glyceryl stearates and monostearates;sorbitan esters, such as spans and TWEENs; polyglyceryl esters, such aspolyglyceryl 4-oleate; fatty alcohol ethoxylates, such as Brij typeemulsifiers; ethoxylated propoxylated block copolymers, such aspoloxamers; polyethylene glycol esters of fatty acids, such as PEG 300linoleic glycerides or Labrafil 2125 CS, PEG 300 oleic glycerides orLabrafil M 1944 CS, PEG 400 caprylic/capric glycerides or Labrasol, andPEG 300 caprylic/capric glycerides or Softigen 767; cremophors, such asCremophor E, polyoxyl 35 castor oil or Cremophor EL, Cremophor EL-P,Cremophor RH 40P, polyoxyl 40 hydrogenated castor oil, Cremophor RH40;polyoxyl 60 hydrogenated castor oil or Cremophor RH 60, glycerolmonocaprylate/caprate, such as Campmul CM 10; polyoxyethylated fattyacids (PEG-stearates, PED-laurates, Brij®), polyoxylated glycerides offatty acid, polyoxylated glycerol fatty acid esters i.e. Solutol HS-15;PEG-ethers (Mirj®), sorbitan derivatives (TWEENs), sorbitan monooleateor Span 20, aromatic compounds (Tritons®), PEG-glycerides (PECEOL™),PEG-PPG (polypropylene glycol) copolymers (PLURONICS including but notlimited to PLURONICS F108, F127, and F68, Poloxamers, Jeffamines),Tetronics, Polyglycerines, PEG-tocopherols, PEG-LICOL 6-oleate;propylene glycol derivatives, sugar and polysaccharide alkyl and acylderivatives (octylsucrose, sucrose stearate, laurolydextran etc.) and/ora mixture thereof; surfactants based on an oleate or laureate ester of apolyalcohol copolymerized with ethylene oxide; Labrasol Gelucire 44/14;polyoxytheylene stearates; saturated polyglycolyzed glycerides; orpoloxamers; all of which are commercially available. Polyoxyethylenesorbitan fatty acid esters can include polysorbates, for example,polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80.Polyoxyethylene stearates can include polyoxyl 6 stearate, polyoxyl 8stearate, polyoxyl 12 stearate and polyoxyl 20 stearate. Saturatedpolyglycolyzed glycerides are, for example, GELUCIRE 44/14 or GELUCIRE™50/13 (Gattefosse, Westwood, N.J., U.S.A.). Poloxamers used hereininclude poloxamer 124 and poloxamer 188.

Surfactants include d-α-tocopheryl polyethylene glycol 1000 succinate(TPGS), polyoxyl 8 stearate (PEG 400 monostearate), polyoxyl 40 stearate(PEG 1750 monostearate) and peppermint oil.

In some variations, the surfactant is polyethoxylated castor oil (e.g.,Cremophor (PEG 35 castor oil)).

In some variations, surfactants having an HLB lower than 10 are used.Such surfactants may optionally be used in combination with othersurfactants as co-surfactants. Examples of some surfactants, mixtures,and other equivalent compositions having an HLB less than or equal to 10are propylene glycols, glyceryl fatty acids, glyceryl fatty acid esters,polyethylene glycol esters, glyceryl glycol esters, polyglycolyzedglycerides and polyoxyethyl steryl ethers. Propylene glycol esters orpartial esters form the composition of commercial products, such asLauroglycol FCC, which contains propylene glycol laureate. Thecommercially available excipient Maisine 35-1 comprises long chain fattyacids, for example glyceryl linoleate. Products, such as Acconon E,which comprise polyoxyethylene stearyl ethers, may also be used.Labrafil M 1944 CS is one example of a surfactant wherein thecomposition contains a mixture of glyceryl glycol esters andpolyethylene glycol esters.

Solubilizing Agents for Rapamycin

Many solubilizing agents may be used for rapamycin, including but notlimited to those in the solubilizing agents section above.

In some variations the solubilizing agent is a surfactant. Nonlimitingexamples of surfactants that may be used for rapamycin include but arenot limited to surfactants with an HLB greater than 10, 11, 12, 13 or14. One nonlimiting example is Cremophor EL. In some variations, thesurfactant is polyethoxylated castor oil (e.g., Cremophor (PEG 35 castoroil)). In some variations, the surfactant may be a polymeric surfactantincluding but not limited to PLURONICS F108, F127, and F68, andTetronics. As noted herein, some solvents may also serve as surfactants.Those of ordinary skill in the art will find it routine to identifywhich solubilizing agents and surfactants may be used for rapamycingiven the teachings herein.

Viscosity Modifying Agents

The liquid formulations described herein may be administered with orfurther comprise a viscosity modifying agent.

One exemplary viscosity modifying agent that may be used is hyaluronicacid. Hyaluronic acid is a glycosaminoglycan. It is made of a repetitivesequence of glucuronic acid and glucosamine. Hyaluronic acid is presentin many tissues and organs of the body, and contributes to the viscosityand consistency of such tissues and organs. Hyaluronic acid is presentin the eye, including the vitreous of the eye, and along with collagencontributes to the viscosity thereof. The liquid formulations describedherein may further comprise or be administered with hyaluronic acid.

Other nonlimiting examples of viscosity modifying agents includepolyalkylene oxides, glycerol, carboxymethyl cellulose, sodium alginate,chitosan, dextran, dextran sulfate and collagen. These viscositymodifying agents can be chemically modified.

Other viscosity modifying agents that may be used include but are notlimited to carrageenan, cellulose gel, colloidal silicon dioxide,gelatin, propylene carbonate, carbonic acid, alginic acid, agar,carboxyvinyl polymers or carbomers and polyacrylamides, acacia, estergum, guar gum, gum arabic, ghatti, gum karaya, tragacanth, terra,pectin, tamarind seed, larch arabinogalactan, alginates, locust bean,xanthan gum, starch, veegum, tragacanth, polyvinyl alcohol, gellan gum,hydrocolloid blends, and povidone. Other viscosity modifying agentsknown in the art can also be used, including but not limited to sodiumcarboxymethyl cellulose, algin, carageenans, galactomannans, hydropropylmethyl cellulose, hydroxypropyl cellulose, polyethylene glycol,polyvinylpyrrolidone, sodium carboxymethyl chitin, sodium carboxymethyldextran, sodium carboxymethyl starch, xanthan gum, and zein.

Other Components of Liquid Formulations

The formulations described herein may further comprise various othercomponents such as stabilizers, for example. Stabilizers that may beused in the formulations described herein include but are not limited toagents that will (1) improve the compatibility of excipients with theencapsulating materials such as gelatin, (2) improve the stability (e.g.prevent crystal growth of a therapeutic agent such as rapamycin) of atherapeutic agent such as rapamycin and/or rapamycin derivatives, and/or(3) improve formulation stability. Note that there is overlap betweencomponents that are stabilizers and those that are solvents,solubilizing agents or surfactants, and the same component can carry outmore than one role.

Stabilizers may be selected from fatty acids, fatty alcohols, alcohols,long chain fatty acid esters, long chain ethers, hydrophilic derivativesof fatty acids, polyvinylpyrrolidones, polyvinylethers, polyvinylalcohols, hydrocarbons, hydrophobic polymers, moisture-absorbingpolymers, and combinations thereof. Amide analogues of the abovestabilizers can also be used. The chosen stabilizer may change thehydrophobicity of the formulation (e.g. oleic acid, waxes), or improvethe mixing of various components in the formulation (e.g. ethanol),control the moisture level in the formula (e.g. PVP), control themobility of the phase (substances with melting points higher than roomtemperature such as long chain fatty acids, alcohols, esters, ethers,amides etc. or mixtures thereof; waxes), and/or improve thecompatibility of the formula with encapsulating materials (e.g. oleicacid or wax). Some of these stabilizers may be used assolvents/co-solvents (e.g. ethanol). Stabilizers may be present insufficient amount to inhibit the therapeutic agent's (such asrapamycin's) crystallization.

Examples of stabilizers include, but are not limited to, saturated,monoenoic, polyenoic, branched, ring-containing, acetylenic,dicarboxylic and functional-group-containing fatty acids such as oleicacid, caprylic acid, capric acid, caproic acid, lauric acid, myristicacid, palmitic acid, stearic acid, behenic acid, linoleic acid,linolenic acid, eicosapentaenoic acid (EPA), DHA; fatty alcohols such asstearyl alcohol, cetyl alcohol, ceteryl alcohol; other alcohols such asethanol, isopropyl alcohol, butanol; long chain fatty acid esters,ethers or amides such as glyceryl stearate, cetyl stearate, oleylethers, stearyl ethers, cetyl ethers, oleyl amides, stearyl amides;hydrophilic derivatives of fatty acids such as polyglyceryl fatty acids,polyethylene glycol fatty acid esters; polyvinylpyrrolidones,polyvinylalcohols (PVAs), waxes, docosahexaenoic acid andde-hydroabietic acid etc.

The formulations described may further contain a gelling agent thatalters the texture of the final formulation through formation of a gel.

The therapeutic agents for use as described herein, such as rapamycin,may be subjected to conventional pharmaceutical operations, such assterilization and compositions containing the therapeutic agent may alsocontain conventional adjuvants, such as preservatives, stabilizers,wetting agents, emulsifiers, buffers etc. The therapeutic agents mayalso be formulated with pharmaceutically acceptable excipients forclinical use to produce a pharmaceutical composition. Formulations forocular administration may be presented as a solution, suspension,particles of solid material, a discrete mass of solid material,incorporated within a polymer matrix, liquid formulations or in anyother form for ocular administration. The therapeutic agents may be usedto prepare a medicament to treat, prevent, inhibit, delay onset, orcause regression of any of the conditions described herein. In somevariations, the therapeutic agents may be used to prepare a medicamentto treat any of the conditions described herein.

A composition containing a therapeutic agent such as rapamycin maycontain one or more adjuvants appropriate for the indicated route ofadministration. Adjuvants with which the therapeutic agent may beadmixed with include but are not limited to lactose, sucrose, starchpowder, cellulose esters of alkanoic acids, stearic acid, talc,magnesium stearate, magnesium oxide, sodium and calcium salts ofphosphoric and sulphuric acids, acacia, gelatin, sodium alginate,polyvinylpyrrolidine, and/or polyvinyl alcohol. When a solubilizedformulation is required the therapeutic agent may be in a solventincluding but not limited to polyethylene glycol of various molecularweights, propylene glycol, carboxymethyl cellulose colloidal solutions,methanol, ethanol, DMSO, corn oil, peanut oil, cottonseed oil, sesameoil, tragacanth gum, and/or various buffers. Other adjuvants and modesof administration are well known in the pharmaceutical art and may beused in the practice of the methods, compositions and liquidformulations described herein. The carrier or diluent may include timedelay material, such as glyceryl monostearate or glyceryl distearatealone or with a wax, or other materials well known in the art. Theformulations for use as described herein may also include gelformulations, erodible and non-erodible polymers, microspheres, andliposomes.

Other adjuvants and excipients that may be used include but are notlimited to C₈-C₁₀ fatty acid esters such as softigen 767, polysorbate80, PLURONICS, Tetronics, Miglyol, and Transcutol.

Additives and diluents normally utilized in the pharmaceutical arts canoptionally be added to the pharmaceutical composition and the liquidformulation. These include thickening, granulating, dispersing,flavoring, sweetening, coloring, and stabilizing agents, including pHstabilizers, other excipients, anti-oxidants (e.g., tocopherol, BHA,BHT, TBHQ, tocopherol acetate, ascorbyl palmitate, ascorbic acid propylgallate, and the like), preservatives (e.g., parabens), and the like.Exemplary preservatives include, but are not limited to, benzylalcohol,ethylalcohol, benzalkonium chloride, phenol, chlorobutanol, and thelike. Some useful antioxidants provide oxygen or peroxide inhibitingagents for the formulation and include, but are not limited to,butylated hydroxytoluene, butylhydroxyanisole, propyl gallate, ascorbicacid palmitate, α-tocopherol, and the like. Thickening agents, such aslecithin, hydroxypropylcellulose, aluminum stearate, and the like, mayimprove the texture of the formulation.

In some variations, the therapeutic agent is rapamycin, and therapamycin is formulated as rapamune in solid or liquid form. In somevariations, the rapamune is formulated as an oral dosage.

In addition, a viscous polymer may be added to the suspension, assistingthe localization and ease of placement and handling. In some uses of theliquid formulation, a pocket in the sclera may be surgically formed toreceive an injection of the liquid formulations. The hydrogel structureof the sclera can act as a rate-controlling membrane. Particles oftherapeutic agent substance for forming a suspension can be produced byknown methods including but not limited to via ball milling, for exampleby using ceramic beads. For example, a Cole Parmer ball mill such asLabmill 8000 may be used with 0.8 mm YTZ ceramic beads available fromTosoh or Norstone Inc.

The formulations may conveniently be presented in unit dosage form andmay be prepared by conventional pharmaceutical techniques. Suchtechniques include the step of bringing into association the therapeuticagent and the pharmaceutical carrier(s) or excipient(s). Theformulations may be prepared by uniformly and intimately bringing intoassociate the active ingredient with liquid carriers or finely dividedsolid carriers or both, and then, if necessary, shaping the product.

In some variations, the formulations described herein are provided inone or more unit dose forms, wherein the unit dose form contains anamount of a liquid formulation described herein that is effective totreat or prevent the disease or condition for which it is beingadministered. In some variations, the formulations described herein areprovided in one or more unit dose forms, wherein the unit dose formcontains an amount of a liquid rapamycin formulation described hereinthat is effective to treat or prevent the disease or condition for whichit is being administered.

In some embodiments, the unit dose form is prepared in the concentrationat which it will be administered. In some variations, the unit dose formis diluted prior to administration to a subject. In some variations, aliquid formulation described herein is diluted in an aqueous mediumprior to administration to a subject. In some variations the aqueousmedium is an isotonic medium. In some variations, a liquid formulationdescribed herein is diluted in an non-aqueous medium prior toadministration to a subject.

In a further aspect, provided herein are kits comprising one or moreunit dose forms as described herein. In some embodiments, the kitcomprises one or more of packaging and instructions for use to treat oneor more diseases or conditions. In some embodiments, the kit comprises adiluent which is not in physical contact with the formulation orpharmaceutical formulation. In some embodiments, the kit comprises anyof one or more unit dose forms described herein in one or more sealedvessels. In some embodiments, the kit comprises any of one or moresterile unit dose forms.

In some variations, the unit dose form is in a container, including butnot limited to a sterile sealed container. In some variations thecontainer is a vial, ampule, or low volume applicator, including but notlimited to a syringe. In some variations, a low-volume applicator ispre-filled with rapamycin for treatment of an ophthalmic disease orcondition, including but not limited to a limus compound for treatmentof age-related macular degeneration. Described herein is a pre-filledlow-volume applicator pre-filled with a formulation comprising atherapeutic agent, including but not limited to rapamycin. In somevariations a low-volume applicator is pre-filled with a solutioncomprising a therapeutic agent, including but not limited to rapamycinand a polyethylene glycol, and optionally further comprises one or moreadditional components including but not limited to ethanol. In somevariations a pre-filled low-volume applicator is pre-filled with asolution comprising about 2% rapamycin, about 94% PEG-400, about 4%ethanol.

Described herein are kits comprising one or more containers. In somevariations a kit comprises one or more low-volume applicators ispre-filled with a formulation described herein comprising a therapeuticagent, including but not limited to formulations comprising rapamycin,formulations comprising rapamycin and a polyethylene glycol, andoptionally further comprises one or more additional components includingbut not limited to ethanol, and formulations in liquid form comprisingabout 2% rapamycin, about 94% PEG-400, about 4% ethanol. In somevariations the kit comprises one or more containers, including but notlimited to pre-filled low-volume applicators, with instructions for itsuse. In a further variation a kit comprises one or more low-volumeapplicators pre-filled with rapamycin, with instructions for its use intreating a disease or condition of the eye. In some variations, thecontainers described herein are in a secondary packaging.

Routes of Administration

The compositions, methods, and liquid formulations described hereindeliver one or more therapeutic agents to a subject, including but notlimited to a human subject.

In some variations, the compositions, methods, and liquid formulationsdescribed herein deliver one or more therapeutic agents to an aqueousmedium of a human subject.

In some variations, the compositions, methods, and liquid formulationsdescribed herein deliver one or more therapeutic agents to an aqueousmedium in or proximal to an area where a disease or condition is to betreated, prevented, inhibited, onset delayed, or regression caused.

In some variations, the compositions, methods, and liquid formulationsdescribed herein deliver one or more therapeutic agents to an eye of asubject, including the macula and the retina choroid tissues, in anamount and for a duration effective to treat, prevent, inhibit, delaythe onset of, or cause the regression of the diseases and conditionsdescribed in the Diseases and Conditions section.

“Retina choroid” and “retina choroid tissues,” as used herein, aresynonymous and refer to the combined retina and choroid tissues of theeye.

As a non-limiting example, the compositions, liquid formulations, andmethods described in herein may be administered to the vitreous, aqueoushumor, sclera, conjunctiva, between the sclera and conjunctiva, theretina choroid tissues, macula, or other area in or proximate to the eyeof a subject, either by direct administration to these tissues or byperiocular routes, in amounts and for a duration effective to treat,prevent, inhibit, delay the onset of, or cause the regression of CNV andwet AMD. The effective amounts and durations may be different for eachof treating, preventing, inhibiting, delaying the onset of, or causingthe regression of CNV and wet AMD, and for each of the different sitesof delivery.

Intravitreal administration is more invasive than some other types ofocular procedures. Because of the potential risks of adverse effects,intravitreal administration may not be optimal for treatment ofrelatively healthy eyes. By contrast, periocular administration, such assubconjunctival administration, is much less invasive than intravitrealadministration. When a therapeutic agent is delivered by a periocularroute, it may be possible to treat patients with healthier eyes thancould be treated using intravitreal administration. In some variations,subconjunctival injection is used to prevent or delay onset of a diseaseor condition of the eye, where the eye of the subject has visual acuityof 20/40 or better.

“Subconjunctival” placement or injection, as used herein, refers toplacement or injection between the sclera and conjunctiva.Subconjunctival is sometimes referred to herein as “sub-conj”administration.

Routes of administration that may be used to administer a liquidformulation include but are not limited to placement of the liquidformulation, for example by injection, into an aqueous medium in thesubject, including but not limited to placement, including but notlimited to by injection, into the eye of a subject, including but notlimited to a human subject. The liquid formulation may be administeredsystemically, including but not limited to the following deliveryroutes: rectal, vaginal, infusion, intramuscular, intraperitoneal,intraarterial, intrathecal, intrabronchial, intracisternal, cutaneous,subcutaneous, intradermal, transdermal, intravenous, intracervical,intraabdominal, intracranial, intraocular, intrapulmonary,intrathoracic, intratracheal, nasal, buccal, sublingual, oral,parenteral, or nebulised or aerosolized using aerosol propellants.

Compositions and liquid formulations comprising therapeutic agent can beadministered directly to the eye using a variety of procedures,including but not limited to procedures in which (1) the therapeuticagent is administered by injection using a syringe and hypodermicneedle, (2) a specially designed device is used to inject thetherapeutic agent, (3) prior to injection of the therapeutic agent, apocket is surgically formed within the sclera to serve as a receptaclefor the therapeutic agent or therapeutic agent composition. For example,in one administration procedure a surgeon forms a pocket within thesclera of the eye followed by injection of a solution or liquidformulation comprising the therapeutic agent into the pocket.

Other administration procedures include, but are not limited toprocedures in which (1) a formulation of the therapeutic agent isinjected through a specially designed curved cannula to place thetherapeutic agent directly against a portion of the eye, (2) acompressed form of the therapeutic agent is placed directly against aportion of the eye, (3) the therapeutic agent is inserted into thesclera by a specially designed injector or inserter, (4) the liquidformulation comprising the therapeutic agent is incorporated within apolymer, (5) a surgeon makes a small conjunctival incision through whichto pass a suture and any therapeutic agent delivery structure so as tosecure the structure adjacent to the sclera, (6) a needle is used forinjection directly into the vitreous of an eye, or into any other sitedescribed.

The liquid formulations described herein may be used directly, forexample, by injection, as an elixir, for topical administrationincluding but not limited to via eye drops, or in hard or soft gelatinor starch capsules. The capsules may be banded to prevent leakage.

In some variations, the liquid formulations described herein may beadministered by topical administration. In some variations, the topicaladministration is ocular topical administration. In some variations, theocular topical administration includes, but is not limited to,administration via eye drops, contacts, punctual plugs, or other oculardevices. In some variations, the liquid formulation is appliedtopically, including topically to the eye, any of about 1, 2, 3, 4, or 5times per day. In some variations, the liquid formulation is appliedtopically, including topically to the eye, about once or less any ofabout every 1, 2, 3, 4, 5, 6, 7, 10, 14, 21, or 28 day(s). In somevariations, the liquid formulation is applied topically, includingtopically to the eye, about once or less a day. In some variations, theliquid formulation is applied topically, including topically to the eye,about once or less every 5 days. In some variations, the liquidformulation is applied topically, including topically to the eye, aboutonce or less of every 10 days.

Delivery by Injection

One method that may be used to deliver the compositions and liquidformulations described herein is delivery by injection. In this methodcompositions and liquid formulations may be injected into a subject,including but not limited to a human subject, or into a position in orproximate to an eye of the subject for delivery to a subject or to theeye of a subject. Injection includes but is not limited to intraocularand periocular injection. Nonlimiting examples of positions that are inor proximate to an eye of a subject are as follows.

Injection of therapeutic agent into the vitreous may provide a highlocal concentration of therapeutic agent in the vitreous and retina.Further, it has been found that in the vitreous the clearance half-livesof drugs increases with molecular weight.

Intracameral injection, or injection into the anterior chamber of theyeye, may also be used. In one example, up to about 100 μl may beinjected intracamerally.

Periocular routes of delivery may deliver therapeutic agent to theretina without some of the risks of intravitreal delivery. Periocularroutes include but are not limited to subconjunctival, subtenon,retrobulbar, peribulbar and posterior juxtascleral delivery. A“periocular” route of administration means placement near or around theeye. For a description of exemplary periocular routes for retinal drugdelivery, see Periocular routes for retinal drug delivery, Raghava etal. (2004), Expert Opin. Drug Deliv. 1(1):99-114, which is incorporatedherein by reference in its entirety.

In some variations the liquid formulations described herein areadministered intraocularly. Intraocular administration includesplacement or injection within the eye, including in the vitreous.

Subconjunctival injection may be by injection of therapeutic agentunderneath the conjunctiva, or between the sclera and conjunctiva. Inone example, up to about 500 μl may be injected subconjunctivally. Asone nonlimiting example, a needle of up to about 25 to about 30 gaugeand about 30 mm long may be used. Local pressure to the subconjunctivalsite of therapeutic agent administration may elevate delivery of thetherapeutic agent to the posterior segment by reducing local choroidalblood flow.

Subtenon injection may be by injection of therapeutic agent into thetenon's capsule around the upper portion of the eye and into the “belly”of the superior rectus muscle. In one example, up to about 4 ml may beinjected subtenon. As one nonlimiting example, a blunt-tipped cannulaabout 2.5 cm long may be used.

Retrobulbar injection refers to injection into the conical compartmentof the four rectus muscles and their intermuscular septa, behind theglobe of the eye. In one example, up to about 5 ml may be injectedretrobulbarly. As one nonlimiting example, a blunt needle of about 25-or about 27-gauge may be used.

Peribulbar injection may be at a location external to the confines ofthe four rectus muscles and their intramuscular septa, i.e., outside ofthe muscle cone. A volume of, for example, up to about 10 ml may beinjected peribulbarly. As one nonlimiting example, a blunt-tippedcannula about 1.25 inches long and about 25-gauge may be used.

Posterior juxtascleral delivery refers to placement of a therapeuticagent near and above the macula, in direct contact with the outersurface of the sclera, and without puncturing the eyeball. In oneexample, up to about 500 ml may be injected posterior juxtasclerally. Asone nonlimiting example, a blunt-tipped curved cannula, speciallydesigned at 56°, is used to place the therapeutic agent in an incisionin the sclera.

In some variations the liquid formulations described herein are injectedintraocularly. Intraocular injection includes injection within the eye.

Sites to which the compositions and liquid formulations may beadministered include but are not limited to the vitreous, aqueous humor,sclera, conjunctiva, between the sclera and conjunctiva, the retinachoroid tissues, macula, or other area in or proximate to the eye of asubject. Methods that may be used for placement of the compositions andliquid formulations include but are not limited to injection.

In one method that may be used, the therapeutic agent is dissolved in ansolvent or solvent mixture and then injected into or proximate to thevitreous, aqueous humor, sclera, conjunctiva, between the sclera andconjunctiva, the retina choroid tissues, macula, other area in orproximate to the eye of a subject, or other medium of a subject,according to any of the procedures mentioned above. In one such methodthat may be used, the therapeutic agent is rapamycin in a liquidformulation.

When the therapeutic agent is rapamycin, the compositions and liquidformulations may be used to deliver or maintain an amount of rapamycinin tissues of the eye, including without limitation retina, choroid, orthe vitreous, which amount is effective to treat AMD. In one nonlimitingexample, it is believed that a liquid formulation delivering rapamycinin an amount capable of providing a concentration of rapamycin of about0.1 pg/ml to about 2 μg/ml in the vitreous may be used for treatment ofwet AMD. In some nonlimiting examples, it is believed that a liquidformulation delivering a concentration of rapamycin of about 0.1 μg/mgto about 1 μg/mg in the retina choroid tissues may be used for treatmentof wet AMD. Other effective concentrations are readily ascertainable bythose of skill in the art based on the teachings described herein.

“Subtenonally administered”, as used herein, means administration to thesubtenon. In some variations, a therapeutic agent or liquid formulationis subtenonally administered by subtenon injection. In some variations,a therapeutic agent or liquid formulation is subtenonally administeredby a means other than subtenon injection.

Method of Preparing Liquid Formulations

One nonlimiting method that may be used for preparing the liquidformulations described herein, including but not limited to liquidformulations comprising rapamycin, is by mixing a solvent and atherapeutic agent together at room temperature or at slightly elevatedtemperature until a solution or suspension is obtained, with optionaluse of a sonicator, and then cooling the formulation. Other componentsincluding but not limited to those described above may then be mixedwith the formulation. Other preparation methods that may be used aredescribed herein including in the examples, and those of skill in theart will be able to select other preparation methods based on theteachings herein.

Extended Delivery of Therapeutic Agents

Described herein are compositions and liquid formulations showing invivo delivery or clearance profiles with one or more of the followingcharacteristics. The delivery or clearance profiles are for clearance ofthe therapeutic agent in vivo after injection of the composition orliquid formulations subconjunctivally or into the vitreous of a rabbiteye. In some variations, the delivery or clearance profiles are forclearance of rapamycin in vivo after injection of the composition orliquid formulations subconjunctivally or into the vitreous of a rabbiteye. The volume of the rabbit vitreous is approximately 30-40% of thevolume of the human vitreous. The amount of therapeutic agent ismeasured using techniques as described in Example 2, but withoutlimitation to the formulation and therapeutic agent described in Example2.

In some variations, the therapeutic agents with the in vivo delivery orclearance profiles described herein include but are not limited to thosedescribed in the Therapeutic Agents section. In some variations thetherapeutic agent is rapamycin. In some variations, the liquidformulations described herein are used to deliver therapeutic agents ina concentration equivalent to rapamycin. The liquid formulationsdescribed herein may comprise any therapeutic agent including but notlimited to those in the Therapeutic Agents section, in a concentrationequivalent to rapamycin including but not limited to thoseconcentrations described herein including in the examples.

“Average percentage in vivo” level means that an average concentrationof therapeutic agent is obtained across multiple rabbit eyes for a giventimepoint, and the average concentration of therapeutic agent at onetimepoint is divided by the average concentration of therapeutic agentat another timepoint. In some variations of the average percentage invivo levels, the therapeutic agent is rapamycin.

The average concentration of a therapeutic agent in the tissue of arabbit eye at a given time after administration of a formulationcontaining the therapeutic agent may be measured according to thefollowing method. Where volumes below 10 μl are to be injected, aHamilton syringe is used.

The liquid formulations are stored at a temperature of 2-8° C. prior touse.

The experimental animals are specific pathogen free (SPF) New ZealandWhite rabbits. A mixed population of about 50% male, about 50% female isused. The rabbits are at least 12 weeks of age, usually at least 14weeks of age, at the time of dosing. The rabbits each weigh at least 2.2kg, usually at least 2.5 kg, at the time of dosing. Prior to the study,the animals are quarantined for at least one week and examined forgeneral health parameters. Any unhealthy animals are not used in thestudy. At least 6 eyes are measured and averaged for a given timepoint.

Housing and sanitation are performed according to standard proceduresused in the industry. The animals are provided approximately 150 gramsof Teklad Certified Hi-Fiber Rabbit Diet daily, and are provided tapwater ad libitum. No contaminants are known to exist in the water and noadditional analysis outside that provided by the local water district isperformed. Environmental Conditions are monitored.

Each animal undergoes a pre-treatment ophthalmic examination (slit lampand opthalmoscopy), performed by a board certified veterinaryophthalmologist. Ocular findings are scored according to the McDonaldand Shadduck scoring system as described in Dermatoxicology, F. N.Marzulli and H. I. Maibach, 1977 “Eye Irritation,” T. O. McDonald and J.A. Shadduck (pages 579-582). Observations are recorded using astandardized data collection sheet. Acceptance criteria for placement onstudy are as follows: scores of <1 for conjunctival congestion andswelling; scores of 0 for all other observation variables.

Gentamicin ophthalmic drops are placed into both eyes of each animaltwice daily on the day prior to dosing, on the day of dosing (Day 1),and on the day after dosing (Day 2). Dosing is performed in two phases,the first including one set of animals and the second including theother animals. Animals are randomized separately into masked treatmentgroups prior to each phase of dosing according to modified Latinsquares. Animals are fasted at least 8 hours prior to injection. Thestart time of the fast and time of injection are recorded.

Animals are weighed and anesthetized with an intravenous injection of aketamine/xylazine cocktail (87 mg/mL ketamine, 13 mg/mL xylazine) at avolume of 0.1-0.2 mL/kg. Both eyes of each animal are prepared forinjection as follows: approximately five minutes prior to injection,eyes are moistened with an ophthalmic Betadine solution. After fiveminutes, the Betadine is washed out of the eyes with sterile saline.Proparacaine hydrochloride 0.5% (1-2 drops) is delivered to each eye.For eyes to be intravitreally injected, 1% Tropicamide (1 drop) isdelivered to each eye.

On Day 1, both eyes of each animal receive an injection of test orcontrol article. Animals in selected groups are dosed a second time onDay 90±1. Dosing is subconjunctival or intravitreal. Actual treatments,injection locations, and dose volumes are masked and revealed at the endof the study.

Subconjunctival injections are given using an insulin syringe and 30gauge×½-inch needle. The bulbar conjunctiva in the dorsotemporalquadrant is elevated using forceps. Test article is injected into thesubconjunctival space.

Intravitreal injections are given using an Insulin syringe and 30gauge×½-inch needle. For each injection, the needle is introducedthrough the ventral-nasal quadrant of the eye, approximately 2-3 mmposterior to the limbus, with the bevel of the needle directed downwardand posteriorly to avoid the lens. Test article is injected in a singlebolus in the vitreous near the retina.

Animals are observed for mortality/morbidity twice daily. An animaldetermined to be moribund is euthanized with an intravenous injection ofcommercial euthanasia solution. Both eyes are removed and stored frozenat −70° C. for possible future evaluation. If an animal is found deadprior to onset of rigor mortis, both eyes are removed and stored frozenat −70° C. for possible future evaluation. Animals found after the onsetof rigor mortis are not necropsied.

Animals are weighed at randomization, on Day 1 prior to dosing, andprior to euthanasia.

Ophthalmic observations (slit lamp and indirect opthalmoscopy) areperformed on all animals on Days 5±1, 30±1, 60±1, 90±1, and at laterdates in some variations. Observations are performed by a boardcertified veterinary ophthalmologist. For animals to be dosed on Day90±1, ophthalmic observations are performed prior to dosing. Ocularfindings are scored according to the McDonald and Shadduck scoringsystem as described in Dermatoxicology, F. N. Marzulli and H. I.Maibach, 1977 “Eye Irritation”, T. O. McDonald and J. A. Shadduck (pages579-582), and observations are recorded using a standardized datacollection sheet.

Whole blood samples (1-3 mL per sample) are collected from each animalprior to necropsy in vacutainer tubes containing EDTA. Each tube isfilled at least ⅔ full and thoroughly mixed for at least 30 seconds.Tubes are stored frozen until shipped on dry ice.

Animals are euthanized with an intravenous injection of commercialeuthanasia solution. Euthanasia is performed according to standardprocedures used in the industry.

For treatment groups dosed intravitreally or subconjunctivally withplacebo, all eyes from each of these groups are placed into Davidsonssolution for approximately 24 hours. Following the 24-hour period, theeyes are transferred to 70% ethanol; these globes are submitted formasked histopathological evaluation by a board certified veterinarypathologist. The time that eyes are placed into Davidsons and the timeof removal are recorded.

For treatment groups dosed intravitreally or subconjunctivally with testarticle, some eyes from each of these groups are frozen at −70° C. andsubmitted for pharmacokinetic analysis. The remaining eyes from each ofthese groups are placed into Davidsons solution for approximately 24hours. Following the 24-hour period, the eyes are transferred to 70%ethanol; these globes are submitted for masked histopathologicalevaluation by a board certified veterinary pathologist. The time thateyes are placed into Davidsons and the time of removal are recorded.

Frozen samples submitted for pharmacokinetic analysis are dissected withdisposable instruments. One set of instruments is used per eye, and thendiscarded. The samples are thawed at room temperature for 1 to 2 minutesto ensure that the frost around the tissue has been removed. The sclerais dissected into 4 quadrants, and the vitreous is removed. If anon-dispersed mass (NDM) is clearly visible within the vitreous, thevitreous is separated into two sections. The section with the NDM isapproximately two-thirds of the vitreous. The section without the NDM isthe portion of the vitreous that is the most distant from the NDM. Theaqueous humor, lens, iris, and cornea are separated. The retina choroidtissue is removed using a forceps and collected for analysis. Theconjunctiva is separated from the sclera.

The various tissue types are collected into separate individualpre-weighed vials which are then capped and weighed. The vials of tissueare stored at −80° C. until analyzed.

The sirolimus content of the retina choroid, sclera, vitreous humor, andwhole anti-coagulated blood is determined by high-pressure liquidchromatography/tandem mass spectroscopy (HPLC/MS/MS) using32-O-desmethoxyrapamycin as an internal standard. Where an NDM wasobserved in the vitreous, the section of the vitreous containing the NDMand the section of the vitreous not containing the NDM are analyzedseparately.

The average concentration of a therapeutic agent over a period of timemeans for representative timepoints over the period of time the averageconcentration at each time point. For example, if the time period is 30days, the average concentration may be measured at 5 day intervals: forthe average concentration at day 5, the average of a number ofmeasurements of concentration at day 5 would be calculated; for theaverage concentration at day 10, the average of a number of measurementsof the concentration at day 10 would be calculated, etc.

In some variations, the liquid formulations described herein may have invivo delivery to the vitreous profiles with the following describedcharacteristics, where the delivery profiles are for delivery oftherapeutic agent in vivo after injection of the liquid formulationbetween the sclera and the conjunctiva of a rabbit eye. One nonlimitingvariation of in vivo delivery to the vitreous profiles is shown in FIG.2.

At day 40 after injection, the average percentage in vivo vitreal levelmay be between about 70% and about 100%, and more usually between about80% and about 90%, relative to the level present at day 20 afterinjection. At day 40 after injection, the average percentage in vivovitreal level may be greater than about 70%, and more usually greaterthan about 80%, relative to the level present at day 20 after injection.

At day 67 after injection, the average percentage in vivo vitreal levelmay be between about 75% and about 115%, and more usually between about85% and about 105%, relative to the level present at day 20 afterinjection. At day 67 after injection, the average percentage in vivovitreal level may be greater than about 75%, and more usually greaterthan about 85%, relative to the level present at day 20 after injection.

At day 90 after injection, the average percentage in vivo vitreal levelmay be between about 20% and about 50%, and more usually between about30% and about 40%, relative to the level present at day 20 afterinjection. At day 90 after injection, the average percentage in vivovitreal level may be greater than about 20%, and more usually greaterthan about 30%, relative to the level present at day 20 after injection.

In some variations, the average percentage in vivo vitreal level has thefollowing characteristics relative to the level present at day 20 afterinjection: at 40 days after injection it is less than about 100%; at 67days after injection it is less than about 115%; and 90 days afterinjection it is less than about 50%.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the vitreous of therabbit eye of at least about 0.01 ng/mL for at least about 30, at leastabout 60, or at least about 90 days after administration of the liquidformulation to the rabbit eyes. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the vitreous of the rabbit eye of at least about0.1 ng/mL for at least about 30, at least about 60, or at least about 90days after administration of the liquid formulation to the rabbit eyes.In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the vitreous of therabbit eye of at least about 1 ng/mL for at least about 30, at leastabout 60, or at least about 90 days after administration of the liquidformulation to the rabbit eyes.

In some variations, the liquid formulations described herein may have invivo delivery to the retina choroid profiles with the followingdescribed characteristics, where the delivery profiles are for deliveryof therapeutic agent in vivo after injection of the liquid formulationbetween the sclera and the conjunctiva of a rabbit eye.

At day 40 after injection, the average percentage in vivo retina choroidlevel may be between about 350% and about 410%, and more usually betweenabout 360% and about 400%, relative to the level present at day 20 afterinjection. At day 40 after injection, the average percentage in vivoretina choroid level may be greater than about 350%, and more usuallygreater than about 360%, relative to the level present at day 20 afterinjection.

At day 67 after injection, the average percentage in vivo retina choroidlevel may be between about 125% and about 165%, and more usually betweenabout 135% and about 155%, relative to the level present at day 20 afterinjection. At day 67 after injection, the average percentage in vivoretina choroid level may be greater than about 125%, and more usuallygreater than about 135%, relative to the level present at day 20 afterinjection.

At day 90 after injection, the average percentage in vivo retina choroidlevel may be between about 10% and about 50%, and more usually betweenabout 20% and about 40%, relative to the level present at day 20 afterinjection. At day 90 after injection, the average percentage in vivoretina choroid level may be greater than about 10%, and more usuallygreater than about 20%, relative to the level present at day 20 afterinjection.

In some variations, the average percentage in vivo retina choroid levelhas the following characteristics relative to the level present at day20 after injection: at 40 days after injection it is less than about410%; at 67 days after injection it is less than about 165%; and 90 daysafter injection it is less than about 50%.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the retina choroidtissues of the rabbit eye of at least about 0.001 ng/mg for at leastabout 30, at least about 60, or at least about 90 days afteradministration of the liquid formulation to the rabbit eyes. In somevariations, the liquid formulation when injected between the sclera andconjunctiva of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye of at least about 0.01 ng/mg for at least about 30, at leastabout 60, or at least about 90 days after administration of the liquidformulation to the rabbit eyes.

In some variations, the level of therapeutic agent present in the retinachoroid first increases, then peaks and decreases. The peak may, forinstance, occur at about day 40 after injection.

In some variations, the liquid formulations described herein may have invivo clearance from the sclera profiles with the following describedcharacteristics, where the clearance profiles are for clearance oftherapeutic agent in vivo after injection of the liquid formulationbetween the sclera and the conjunctiva of a rabbit eye. Where injectionis between the sclera and the conjunctiva, the scleral level is thoughtto include the injected liquid formulation.

At day 40 after injection, the average percentage in vivo scleral levelmay be between about 150% and about 230%, and more usually between about170% and about 210%, relative to the level present at day 20 afterinjection. At day 40 after injection, the average percentage in vivoscleral level may be greater than about 150%, and more usually greaterthan about 170%, relative to the level present at day 20 afterinjection.

At day 67 after injection, the average percentage in vivo scleral levelmay be between about 30% and about 70%, and more usually between about40% and about 60%, relative to the level present at day 20 afterinjection. At day 67 after injection, the average percentage in vivoscleral level may be greater than about 30%, and more usually greaterthan about 40%, relative to the level present at day 20 after injection.

At day 90 after injection, the average percentage in vivo scleral levelmay be between about 110% and about 160%, and more usually between about125% and about 145%, relative to the level present at day 20 afterinjection. At day 90 after injection, the average percentage in vivoscleral level may be greater than about 110%, and more usually greaterthan about 125%, relative to the level present at day 20 afterinjection.

In some variations, the average percentage in vivo scleral level has thefollowing characteristics relative to the level present at day 20 afterinjection: at 40 days after injection it is less than about 230%; at 67days after injection it is less than about 70%; and 90 days afterinjection it is less than about 160%.

In some variations, the level of therapeutic agent present in the sclerafirst increases, then peaks and decreases. The peak may, for instance,occur at about day 40 after injection.

In some variations, the liquid formulations described herein may have invivo delivery to the vitreous profiles with the following describedcharacteristics, where the delivery profiles are for delivery oftherapeutic agent in vivo after injection of the liquid formulationbetween the sclera and the conjunctiva of a rabbit eye.

At day 14 after injection, the average percentage in vivo vitreal levelmay be between about 1350% and about 1650%, and more usually betweenabout 1450% and about 1550%, relative to the level present at day 2after injection. At day 14 after injection, the average percentage invivo vitreal level may be greater than about 1350%, and more usuallygreater than about 1450%, relative to the level present at day 2 afterinjection.

At day 35 after injection, the average percentage in vivo vitreal levelmay be between about 200% and about 300%, and more usually between about225% and about 275%, relative to the level present at day 2 afterinjection. At day 35 after injection, the average percentage in vivovitreal level may be greater than about 200%, and more usually greaterthan about 225%, relative to the level present at day 2 after injection.

At day 62 after injection, the average percentage in vivo vitreal levelmay be between about 100% and about 160%, and more usually between about115% and about 145%, relative to the level present at day 2 afterinjection. At day 62 after injection, the average percentage in vivovitreal level may be greater than about 100%, and more usually greaterthan about 115%, relative to the level present at day 2 after injection.

At day 85 after injection, the average percentage in vivo vitreal levelmay be between about 5% and about 30%, and more usually between about10% and about 25%, relative to the level present at day 2 afterinjection. At day 85 after injection, the average percentage in vivovitreal level may be greater than about 5%, and more usually greaterthan about 10%, relative to the level present at day 2 after injection.

In some variations, the average percentage in vivo vitreal level has thefollowing characteristics relative to the level present at day 2 afterinjection: at 14 days after injection it is less than about 1600%; at 35days after injection it is less than about 300%; at 62 days afterinjection it is less than about 160% and 85 days after injection it isless than about 30%.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the vitreous of therabbit eye of at least about 0.01 ng/mL for at least about 30, at leastabout 60, or at least about 85 days after administration of the liquidformulation to the rabbit eyes. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the vitreous of the rabbit eye of at least about0.1 ng/mL for at least about 30, at least about 60, or at least about 85days after administration of the liquid formulation to the rabbit eyes.In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the vitreous of therabbit eye of at least about 1 ng/mL for at least about 30, or at leastabout 60 days after administration of the liquid formulation to therabbit eyes.

In some variations, the level of therapeutic agent present in thevitreous first increases, then peaks and decreases. The peak may, forinstance, occur at about day 14 after injection.

In some variations, the liquid formulations described herein may have invivo delivery to the retina choroid profiles with the followingdescribed characteristics, where the delivery profiles are for deliveryof therapeutic agent in vivo after injection of the liquid formulationbetween the sclera and the conjunctiva of a rabbit eye.

At day 35 after injection, the average percentage in vivo retina choroidlevel may be between about 320% and about 400%, and more usually betweenabout 340% and about 380%, relative to the level present at day 14 afterinjection. At day 35 after injection, the average percentage in vivoretina choroid level may be greater than about 320%, and more usuallygreater than about 340%, relative to the level present at day 14 afterinjection.

At day 62 after injection, the average percentage in vivo retina choroidlevel may be between about 3% and about 25%, and more usually betweenabout 6% and about 20%, relative to the level present at day 14 afterinjection. At day 62 after injection, the average percentage in vivoretina choroid level may be greater than about 3%, and more usuallygreater than about 6%, relative to the level present at day 14 afterinjection.

At day 85 after injection, the average percentage in vivo retina choroidlevel may be between about 0.1% and about 6%, and more usually betweenabout 0.5% and about 4%, relative to the level present at day 14 afterinjection. At day 85 after injection, the average percentage in vivoretina choroid level may be greater than about 0.1%, and more usuallygreater than about 0.5%, relative to the level present at day 14 afterinjection.

In some variations, the average percentage in vivo retina choroid levelhas the following characteristics relative to the level present at day14 after injection: at 35 days after injection it is less than about400%; at 62 days after injection it is less than about 25%; and 85 daysafter injection it is less than about 6%.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the retina choroidtissues of the rabbit eye of at least about 0.001 ng/mg for at leastabout 30, at least about 60, or at least about 85 days afteradministration of the liquid formulation to the rabbit eyes. In somevariations, the liquid formulation when injected between the sclera andconjunctiva of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye of at least about 0.01 ng/mg for at least about 30, at leastabout 60, or at least about 85 days after administration of the liquidformulation to the rabbit eyes.

In some variations, the liquid formulations described herein may have invivo clearance from the sclera profiles with the following describedcharacteristics, where the clearance profiles are for clearance oftherapeutic agent in vivo after injection of the liquid formulationbetween the sclera and the conjunctiva of a rabbit eye. For injectionbetween the sclera and conjunctiva, the scleral level is thought toinclude the injected liquid formulation.

At day 35 after injection, the average percentage in vivo scleral levelmay be between about 0.1% and about 0.7%, and more usually between about0.2% and about 0.6%, relative to the level present at day 14 afterinjection. At day 35 after injection, the average percentage in vivoscleral level may be greater than about 0.1%, and more usually greaterthan about 0.2%, relative to the level present at day 14 afterinjection.

At day 62 after injection, the average percentage in vivo scleral levelmay be between about 0.05% and about 0.35%, and more usually betweenabout 0.07% and about 0.3%, relative to the level present at day 14after injection. At day 62 after injection, the average percentage invivo scleral level may be greater than about 0.05%, and more usuallygreater than about 0.07%, relative to the level present at day 14 afterinjection.

At day 85 after injection, the average percentage in vivo scleral levelmay be between about 0.1% and about 0.9%, and more usually between about0.3% and about 0.7%, relative to the level present at day 14 afterinjection. At day 85 after injection, the average percentage in vivoscleral level may be greater than about 0.1%, and more usually greaterthan about 0.3%, relative to the level present at day 14 afterinjection.

In some variations, the average percentage in vivo scleral level has thefollowing characteristics relative to the level present at day 14 afterinjection: at 35 days after injection it is less than about 0.7%; at 62days after injection it is less than about 0.35%; and 85 days afterinjection it is less than about 0.9%.

In some variations, the liquid formulations described herein may have invivo clearance from the vitreous profiles with the following describedcharacteristics, where the clearance profiles are for clearance oftherapeutic agent in vivo after injection of the liquid formulation intothe vitreous of a rabbit eye. Where injection is into the vitreous, themeasured vitreous level is thought to include the injected formulation.

At day 35 after injection, the average percentage in vivo vitreal levelmay be between about 1% and about 40%, and more usually between about 1%and about 10%, relative to the level present at day 14 after injection.At day 35 after injection, the average percentage in vivo vitreal levelmay be greater than about 1% relative to the level present at day 14after injection.

At day 62 after injection, the average percentage in vivo vitreal levelmay be between about 1% and about 40%, and more usually between about 5%and about 25%, relative to the level present at day 14 after injection.At day 62 after injection, the average percentage in vivo vitreal levelmay be greater than about 1% relative to the level present at day 14after injection, and more usually greater than about 5% relative to thelevel present at day 14 after injection.

At day 90 after injection, the average percentage in vivo vitreal levelmay be between about 1% and about 40%, and more usually between about10% and about 30%, relative to the level present at day 14 afterinjection. At day 90 after injection, the average percentage in vivovitreal level may be greater than about 1% relative to the level presentat day 14 after injection, and more usually greater than about 10%relative to the level present at day 14 after injection.

In some variations, the level of therapeutic agent present in thevitreous first increases, then peaks and decreases. The peak may, forinstance, occur at about day 14 after injection.

In some variations, the liquid formulations described herein may have invivo delivery to the retina choroid profiles with the followingdescribed characteristics, where the delivery profiles are for deliveryof therapeutic agent in vivo after injection of the liquid formulationinto the vitreous of a rabbit eye.

At day 35 after injection, the average percentage in vivo retina choroidlevel may be between about 3400% and about 5100%, and more usuallybetween about 3750% and about 4750%, relative to the level present atday 14 after injection. At day 35 after injection, the averagepercentage in vivo retina choroid level may be greater than about 3400%,and more usually greater than about 3750%, relative to the level presentat day 14 after injection.

At day 62 after injection, the average percentage in vivo retina choroidlevel may be between about 0.1% and about 5%, and more usually betweenabout 1% and about 3%, relative to the level present at day 14 afterinjection. At day 62 after injection, the average percentage in vivoretina choroid level may be greater than about 0.1%, and more usuallygreater than about 1%, relative to the level present at day 14 afterinjection.

At day 90 after injection, the average percentage in vivo retina choroidlevel may be between about 10% and about 50%, and more usually betweenabout 20% and about 40%, relative to the level present at day 14 afterinjection. At day 90 after injection, the average percentage in vivoretina choroid level may be greater than about 10%, and more usuallygreater than about 20%, relative to the level present at day 14 afterinjection.

In some variations, the average percentage in vivo retina choroid levelhas the following characteristics relative to the level present at day14 after injection: at 35 days after injection it is less than about5100%; at 62 days after injection it is less than about 5%; and 90 daysafter injection it is less than about 50%.

In some variations, the liquid formulations described herein may have invivo delivery to the sclera profiles with the following describedcharacteristics, where the delivery profiles are for delivery oftherapeutic agent in vivo after injection of the liquid formulation intothe vitreous of a rabbit eye.

At day 35 after injection, the average percentage in vivo scleral levelmay be between about 1700% and about 2600%, and more usually betweenabout 1900% and about 2400%, relative to the level present at day 14after injection. At day 35 after injection, the average percentage invivo scleral level may be greater than about 1700%, and more usuallygreater than about 1900%, relative to the level present at day 14 afterinjection.

At day 62 after injection, the average percentage in vivo scleral levelmay be between about 120% and about 180%, and more usually between about140% and about 160%, relative to the level present at day 14 afterinjection. At day 62 after injection, the average percentage in vivoscleral level may be greater than about 120%, and more usually greaterthan about 140%, relative to the level present at day 14 afterinjection.

At day 90 after injection, the average percentage in vivo scleral levelmay be between about 95% and about 155%, and more usually between about115% and about 135%, relative to the level present at day 14 afterinjection. At day 90 after injection, the average percentage in vivoscleral level may be greater than about 95%, and more usually greaterthan about 115%, relative to the level present at day 14 afterinjection.

In some variations, the average percentage in vivo scleral level has thefollowing characteristics relative to the level present at day 14 afterinjection: at 35 days after injection it is less than about 2600%; at 62days after injection it is less than about 180%; and 90 days afterinjection it is less than about 155%.

In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the sclera of the rabbit eye of atleast about 0.001 ng/mg for at least about 30, at least about 60, or atleast about 90 days after administration of the liquid formulation tothe rabbit eyes. In some variations, the liquid formulation wheninjected into the vitreous of a rabbit eye delivers therapeutic agentgiving an average concentration of therapeutic agent in the sclera ofthe rabbit eye of at least about 0.01 ng/mg for at least about 30, atleast about 60, or at least about 90 days after administration of theliquid formulation to the rabbit eyes. In some variations, the liquidformulation when injected into the vitreous of a rabbit eye deliverstherapeutic agent giving an average concentration of therapeutic agentin the sclera of the rabbit eye of at least about 0.1 ng/mg for at leastabout 30, at least about 60, or at least about 90 days afteradministration of the liquid formulation to the rabbit eyes.

In some variations, the level of therapeutic agent present in thevitreous first increases, then peaks and decreases. The peak may, forinstance, occur at about day 35 after injection.

In some variations, in situ gelling liquid formulations described hereinmay have in vivo delivery to the vitreous profiles with the followingdescribed characteristics, where the delivery profiles are for deliveryof therapeutic agent in vivo after injection of the liquid formulationbetween the sclera and the conjunctiva of a rabbit eye.

At day 32 after injection, the average percentage in vivo vitreal levelmay be between about 25% and about 85%, and more usually between about45% and about 65%, relative to the level present at day 7 afterinjection. At day 40 after injection, the average percentage in vivovitreal level may be greater than about 25%, and more usually greaterthan about 45%, relative to the level present at day 7 after injection.

At day 45 after injection, the average percentage in vivo vitreal levelmay be between about 2% and about 50%, and more usually between about 8%and about 20%, relative to the level present at day 7 after injection.At day 67 after injection, the average percentage in vivo vitreal levelmay be greater than about 2%, and more usually greater than about 5%,relative to the level present at day 7 after injection.

At day 90 after injection, the average percentage in vivo vitreal levelmay be between about 40% and about 100%, and more usually between about60% and about 80%, relative to the level present at day 7 afterinjection. At day 90 after injection, the average percentage in vivovitreal level may be greater than about 40%, and more usually greaterthan about 60%, relative to the level present at day 7 after injection.

In some variations, the average percentage in vivo vitreal level has thefollowing characteristics relative to the level present at day 7 afterinjection: at 32 days after injection it is less than about 80%; at 45days after injection it is less than about 30%; and 90 days afterinjection it is less than about 100%.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the vitreous of therabbit eye of at least about 0.1 μg/mL for at least about 30, at leastabout 60, or at least about 90 days after administration of the liquidformulation to the rabbit eye. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the vitreous of the rabbit eye of at least about0.01 ng/mL for at least about 30, at least about 60, or at least about90 days after administration of the liquid formulation to the rabbiteye. In some variations, the liquid formulation when injected betweenthe sclera and conjunctiva of a rabbit eye delivers therapeutic agentgiving an average concentration of therapeutic agent in the vitreous ofthe rabbit eye of at least about 0.1 ng/mL for at least about 30, atleast about 60, or at least about 90 days after administration of theliquid formulation to the rabbit eye. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the vitreous of the rabbit eye of at least about 1ng/mL for at least about 30, at least about 60, or at least about 90days after administration of the liquid formulation to the rabbit eye.In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the vitreous of therabbit eye of at least about 10 ng/mL for at least about 30, at leastabout 60, or at least about 90 days after administration of the liquidformulation to the rabbit eye.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the vitreous of therabbit eye of at least 0.001 ng/mL for at least 30, at least 60, atleast 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the vitreous of the rabbit eye of at least 0.01ng/mL for at least 30, at least 60, at least 90, or at least 120 daysafter administration of the liquid formulation to the rabbit eyes. Insome variations, the liquid formulation when injected between the scleraand conjunctiva of a rabbit eye delivers therapeutic agent giving anaverage concentration of therapeutic agent in the vitreous of the rabbiteye of at least 0.1 ng/mL for at least 30, at least 60, at least 90, orat least 120 days after administration of the liquid formulation to therabbit eyes. In some variations, the liquid formulation when injectedbetween the sclera and conjunctiva of a rabbit eye delivers therapeuticagent giving an average concentration of therapeutic agent in thevitreous of the rabbit eye of at least 0.5 ng/mL for at least 30, atleast 60, at least 90, or at least 120 days after administration of theliquid formulation to the rabbit eyes.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the vitreous of therabbit eye of between 0.001 ng/mL and 10.0 ng/mL for at least 30, atleast 60, at least 90, or at least 120 days after administration of theliquid formulation to the rabbit eyes. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the vitreous of the rabbit eye of between 0.01ng/mL and 10 ng/mL for at least 30, at least 60, at least 90, or atleast 120 days after administration of the liquid formulation to therabbit eyes. In some variations, the liquid formulation when injectedbetween the sclera and conjunctiva of a rabbit eye delivers therapeuticagent giving an average concentration of therapeutic agent in thevitreous of the rabbit eye of between 0.1 ng/mL and 10 ng/mL for atleast 30, at least 60, at least 90, or at least 120 days afteradministration of the liquid formulation to the rabbit eyes.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the vitreous of therabbit eye of between 0.5 ng/mL and 10.0 ng/mL for at least 30, at least60, at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givinga ratio of a maximum average concentration of therapeutic agent in thevitreous of a rabbit eye to a minimum average concentration oftherapeutic agent in the vitreous of a rabbit eye less than 100 for days30 to at least 60, at least 90, or at least 120 days afteradministration of the liquid formulation to the rabbit eyes. In somevariations, the liquid formulation when injected between the sclera andconjunctiva of a rabbit eye delivers therapeutic agent giving a ratio ofa maximum average concentration of therapeutic agent in the vitreous ofa rabbit eye to a minimum average concentration of therapeutic agent inthe vitreous of a rabbit eye less than 50 for days 30 to at least 60, atleast 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving a ratio of a maximum averageconcentration of therapeutic agent in the vitreous of a rabbit eye to aminimum average concentration of therapeutic agent in the vitreous of arabbit eye less than 10 for days 30 to at least 60, at least 90, or atleast 120 days after administration of the liquid formulation to therabbit eyes. In some variations, the liquid formulation when injectedbetween the sclera and conjunctiva of a rabbit eye delivers therapeuticagent giving a ratio of a maximum average concentration of therapeuticagent in the vitreous of a rabbit eye to a minimum average concentrationof therapeutic agent in the vitreous of a rabbit eye less than 5 fordays 30 to at least 60, at least 90, or at least 120 days afteradministration of the liquid formulation to the rabbit eyes.

“Approximately constant,” as used herein, means that the average leveldoes not vary by more than one order of magnitude over the extendedperiod of time, i.e., the difference between the maximum and minimum isless than a 10-fold difference for measurements of the averageconcentration at times in the relevant period of time.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the vitreous of arabbit eye that is approximately constant at a value greater than 0.001ng/mL for days 30 to at least 60, at least 90, or at least 120 daysafter administration of the solution to the rabbit eyes. In somevariations, the liquid formulation when injected between the sclera andconjunctiva of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the vitreous of a rabbit eye thatis approximately constant at a value greater than 0.01 ng/mL for days 30to at least 60, at least 90, or at least 120 days after administrationof the liquid formulation to the rabbit eyes. In some variations, theliquid formulation when injected between the sclera and conjunctiva of arabbit eye delivers therapeutic agent giving an average concentration oftherapeutic agent in the vitreous of a rabbit eye that is approximatelyconstant at a value greater than 0.1 ng/mL for days 30 to at least 60,at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the vitreous of a rabbit eye that is approximatelyconstant at a value of 1.0 ng/mL for days 30 to at least 60, at least90, or at least 120 days after administration of the liquid formulationto the rabbit eyes.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the retina choroidtissues of the rabbit eye of at least 0.001 ng/mg for at least 30, atleast 60, at least 90, or at least 120 days after administration of theliquid formulation to the rabbit eyes. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the retina choroid tissues of the rabbit eye of atleast 0.005 ng/mg for at least 30, at least 60, at least 90, or at least120 days after administration of the liquid formulation to the rabbiteyes. In some variations, the liquid formulation when injected betweenthe sclera and conjunctiva of a rabbit eye delivers therapeutic agentgiving an average concentration of therapeutic agent in the retinachoroid tissues of the rabbit eye of at least 0.01 ng/mg for at least30, at least 60, at least 90, or at least 120 days after administrationof the liquid formulation to the rabbit eyes.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the retina choroidtissues of the rabbit eye of between 0.001 ng/mg and 1.0 ng/mg for atleast 30, at least 60, at least 90, or at least 120 days afteradministration of the liquid formulation to the rabbit eyes. In somevariations, the liquid formulation when injected between the sclera andconjunctiva of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye of between 0.001 ng/mg and 0.50 ng/mg for at least 30, atleast 60, at least 90, or at least 120 days after administration of theliquid formulation to the rabbit eyes. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the retina choroid tissues of the rabbit eye ofbetween 0.001 ng/mg and 0.15 ng/mg for at least 30, at least 60, atleast 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the retina choroid tissues of the rabbit eye ofbetween 0.001 ng/mg and 0.1 ng/mg for at least 30, at least 60, at least90, or at least 120 days after administration of the liquid formulationto the rabbit eyes.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the retina choroidtissues of the rabbit eye of between 0.005 ng/mg and 1.0 ng/mg for atleast 30, at least 60, at least 90, or at least 120 days afteradministration of the liquid formulation to the rabbit eyes. In somevariations, the liquid formulation when injected between the sclera andconjunctiva of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye of between 0.005 ng/mg and 0.50 ng/mg for at least 30, atleast 60, at least 90, or at least 120 days after administration of theliquid formulation to the rabbit eyes. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the retina choroid tissues of the rabbit eye ofbetween 0.005 ng/mg and 0.15 ng/mg for at least 30, at least 60, atleast 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the retina choroid tissues of the rabbit eye ofbetween 0.005 ng/mg and 0.1 ng/mg for at least 30, at least 60, at least90, or at least 120 days after administration of the liquid formulationto the rabbit eyes.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the retina choroidtissues of the rabbit eye of between 0.01 ng/mg and 1.0 ng/mg for atleast 30, at least 60, at least 90, or at least 120 days afteradministration of the liquid formulation to the rabbit eyes. In somevariations, the liquid formulation when injected between the sclera andconjunctiva of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye of between 0.01 ng/mg and 0.50 ng/mg for at least 30, atleast 60, at least 90, or at least 120 days after administration of theliquid formulation to the rabbit eyes. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the retina choroid tissues of the rabbit eye ofbetween 0.01 ng/mg and 0.15 ng/mg for at least 30, at least 60, at least90, or at least 120 days after administration of the liquid formulationto the rabbit eyes. In some variations, the liquid formulation wheninjected between the sclera and conjunctiva of a rabbit eye deliverstherapeutic agent giving an average concentration of therapeutic agentin the retina choroid tissues of the rabbit eye of between 0.01 ng/mgand 0.1 ng/mg for at least 30, at least 60, at least 90, or at least 120days after administration of the liquid formulation to the rabbit eyes.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givinga ratio of a maximum average concentration of therapeutic agent in theretina choroid tissues of a rabbit eye to a minimum averageconcentration of therapeutic agent in the retina choroid tissues of arabbit eye less than 100 for days 30 to at least 60, at least 90, or atleast 120 days after administration of the liquid formulation to therabbit eyes. In some variations, the liquid formulation when injectedbetween the sclera and conjunctiva of a rabbit eye delivers therapeuticagent giving a ratio of a maximum average concentration of therapeuticagent in the retina choroid tissues of a rabbit eye to a minimum averageconcentration of therapeutic agent in the retina choroid tissues of arabbit eye less than 50 for days 30 to at least 60, at least 90, or atleast 120 days after administration of the liquid formulation to therabbit eyes. In some variations, the liquid formulation when injectedbetween the sclera and conjunctiva of a rabbit eye delivers therapeuticagent giving a ratio of a maximum average concentration of therapeuticagent in the retina choroid tissues of a rabbit eye to a minimum averageconcentration of therapeutic agent in the retina choroid tissues of arabbit eye less than 10 for days 30 to at least 60, at least 90, or atleast 120 days after administration of the liquid formulation to therabbit eyes. In some variations, the liquid formulation when injectedbetween the sclera and conjunctiva of a rabbit eye delivers therapeuticagent giving a ratio of a maximum average concentration of therapeuticagent in the retina choroid tissues of a rabbit eye to a minimum averageconcentration of therapeutic agent in the retina choroid tissues of arabbit eye less than 5 for days 30 to at least 60, at least 90, or atleast 120 days after administration of the liquid formulation to therabbit eyes.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the retina choroidtissues of a rabbit eye that is approximately constant at a valuegreater than 0.001 ng/mg for days 30 to at least 60, at least 90, or atleast 120 days after administration of the liquid formulation to therabbit eyes. In some variations, the liquid formulation when injectedbetween the sclera and conjunctiva of a rabbit eye delivers therapeuticagent giving an average concentration of therapeutic agent in the retinachoroid tissues of a rabbit eye that is approximately constant at avalue greater than 0.005 ng/mg for days 30 to at least 60, at least 90,or at least 120 days after administration of the liquid formulation tothe rabbit eyes. In some variations, the liquid formulation wheninjected between the sclera and conjunctiva of a rabbit eye deliverstherapeutic agent giving an average concentration of therapeutic agentin the retina choroid tissues of a rabbit eye that is approximatelyconstant at a value greater than 0.01 ng/mg for days 30 to at least 60,at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes.

In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the vitreous of the rabbit eye ofat least 100 ng/mL for at least 30, at least 60, at least 90, or atleast 120 days after administration of the liquid formulation to therabbit eyes. In some variations, the liquid formulation when injectedinto the vitreous of a rabbit eye delivers therapeutic agent giving anaverage concentration of therapeutic agent in the vitreous of the rabbiteye of at least 1000 ng/mL for at least 30, at least 60, at least 90, orat least 120 days after administration of the liquid formulation to therabbit eyes. In some variations, the liquid formulation when injectedinto the vitreous of a rabbit eye delivers therapeutic agent giving anaverage concentration of therapeutic agent in the vitreous of the rabbiteye of at least 10,000 ng/mL for at least 30, at least 60, at least 90,or at least 120 days after administration of the liquid formulation tothe rabbit eyes.

In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the vitreous of the rabbit eyebetween 100 ng/mL and 100,000 ng/mL for day 30 to at least 60, at least90, or at least 120 days after administration of the liquid formulationto the rabbit eyes. In some variations, the liquid formulation wheninjected into the vitreous of a rabbit eye delivers therapeutic agentgiving an average concentration of therapeutic agent in the vitreous ofthe rabbit eye between 100 ng/mL and 50,000 ng/mL for day 30 to at least60, at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes.

In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the vitreous of the rabbit eyebetween 1000 ng/mL and 100,000 ng/mL for day 30 to at least 60, at least90, or at least 120 days after administration of the liquid formulationto the rabbit eyes. In some variations, the liquid formulation wheninjected into the vitreous of a rabbit eye delivers therapeutic agentgiving an average concentration of therapeutic agent in the vitreous ofthe rabbit eye between 1000 ng/mL and 50,000 ng/mL for day 30 to atleast 60, at least 90, or at least 120 days after administration of theliquid formulation to the rabbit eyes.

In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving a ratio of amaximum average concentration of therapeutic agent in the vitreous ofthe rabbit eye to a minimum average concentration of therapeutic agentin the vitreous of the rabbit eye less than 100 for days 30 to at least60, at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes. In some variations, the liquidformulation when injected into the vitreous of a rabbit eye deliverstherapeutic agent giving a ratio of a maximum average concentration oftherapeutic agent in the vitreous of the rabbit eye to a minimum averageconcentration of therapeutic agent in the vitreous of the rabbit eyeless than 50 for days 30 to at least 60, at least 90, or at least 120days after administration of the liquid formulation to the rabbit eyes.In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving a ratio of amaximum average concentration of therapeutic agent in the vitreous ofthe rabbit eye to a minimum average concentration of therapeutic agentin the vitreous of the rabbit eye less than 10 for days 30 to at least60, at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes.

In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the vitreous of the rabbit eyethat is approximately constant at a value greater than 100 ng/mL fordays 30 to at least 60, at least 90, or at least 120 days afteradministration of the liquid formulation to the rabbit eyes. In somevariations, the liquid formulation when injected into the vitreous of arabbit eye delivers therapeutic agent giving an average concentration oftherapeutic agent in the vitreous of the rabbit eye that isapproximately constant at a value greater than 1000 ng/mL for days 30 toat least 60, at least 90, or at least 120 days after administration ofthe liquid formulation to the rabbit eyes. In some variations, theliquid formulation when injected into the vitreous of a rabbit eyedelivers therapeutic agent giving an average concentration oftherapeutic agent in the vitreous of the rabbit eye that isapproximately constant at a value greater than 10,000 ng/mL for days 30to at least 60, at least 90, or at least 120 days after administrationof the liquid formulation to the rabbit eyes.

In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye of at least 0.001 ng/mg for at least 30, at least 60, atleast 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes. In some variations, the liquidformulation when injected into the vitreous of a rabbit eye deliverstherapeutic agent giving an average concentration of therapeutic agentin the retina choroid tissues of the rabbit eye of at least 0.01 ng/mgfor at least 30, at least 60, at least 90, or at least 120 days afteradministration of the liquid formulation to the rabbit eyes. In somevariations, the liquid formulation when injected into the vitreous of arabbit eye delivers therapeutic agent giving an average concentration oftherapeutic agent in the retina choroid tissues of the rabbit eye of atleast 0.05 ng/mg for at least 30, at least 60, at least 90, or at least120 days after administration of the liquid formulation to the rabbiteyes. In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye of at least 0.10 ng/mg for at least 30, at least 60, at least90, or at least 120 days after administration of the liquid formulationto the rabbit eyes.

In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye between 0.001 ng/mg and 10.00 ng/mg for at least 30, at least60, at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes. In some variations, the liquidformulation when injected into the vitreous of a rabbit eye deliverstherapeutic agent giving an average concentration of therapeutic agentin the retina choroid tissues of the rabbit eye between 0.001 ng/mg and5.00 ng/mg for at least 30, at least 60, at least 90, or at least 120days after administration of the liquid formulation to the rabbit eyes.In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye between 0.001 ng/mg and 1.00 ng/mg for at least 30, at least60, at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes.

In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye between 0.01 ng/mg and 10.00 ng/mg for at least 30, at least60, at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes. In some variations, the liquidformulation when injected into the vitreous of a rabbit eye deliverstherapeutic agent giving an average concentration of therapeutic agentin the retina choroid tissues of the rabbit eye between 0.01 ng/mg and5.00 ng/mg for at least 30, at least 60, at least 90, or at least 120days after administration of the liquid formulation to the rabbit eyes.In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye between 0.01 ng/mg and 1.00 ng/mg for at least 30, at least60, at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes.

In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye between 0.05 ng/mg and 10.00 ng/mg for at least 30, at least60, at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes. In some variations, the liquidformulation when injected into the vitreous of a rabbit eye deliverstherapeutic agent giving an average concentration of therapeutic agentin the retina choroid tissues of the rabbit eye between 0.05 ng/mg and5.00 ng/mg for at least 30, at least 60, at least 90, or at least 120days after administration of the liquid formulation to the rabbit eyes.In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye between 0.05 ng/mg and 1.00 ng/mg for at least 30, at least60, at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes.

In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye between 0.10 ng/mg and 10.00 ng/mg for at least 30, at least60, at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes. In some variations, the liquidformulation when injected into the vitreous of a rabbit eye deliverstherapeutic agent giving an average concentration of therapeutic agentin the retina choroid tissues of the rabbit eye between 0.10 ng/mg and5.00 ng/mg for at least 30, at least 60, at least 90, or at least 120days after administration of the liquid formulation to the rabbit eyes.In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the retina choroid tissues of therabbit eye between 0.10 ng/mg and 1.00 ng/mg for at least 30, at least60, at least 90, or at least 120 days after administration of the liquidformulation to the rabbit eyes.

In some variations, the liquid formulation when injected into thevitreous of a rabbit eye delivers therapeutic agent giving a ratio of amaximum average concentration of therapeutic agent in the retina choroidtissues of the rabbit eye to a minimum average concentration oftherapeutic agent in the retina choroid tissues of the rabbit eye lessthan 100 for days 30 to at least 60, at least 90, or at least 120 daysafter administration of the liquid formulation to the rabbit eyes. Insome variations, the liquid formulation when injected into the vitreousof a rabbit eye delivers therapeutic agent giving a ratio of a maximumaverage concentration of therapeutic agent in the retina choroid tissuesof the rabbit eye to a minimum average concentration of therapeuticagent in the retina choroid tissues of the rabbit eye less than 50 fordays 30 to at least 60, at least 90, or at least 120 days afteradministration of the liquid formulation to the rabbit eyes.

In some variations, in situ gelling liquid formulations described hereinmay have in vivo delivery to the retina choroid tissue profiles with thefollowing described characteristics, where the delivery profiles are fordelivery of therapeutic agent in vivo after injection of the liquidformulation between the sclera and the conjunctiva of a rabbit eye.

At day 32 after injection, the percentage in vivo vitreal level may bebetween about 20% and about 80%, and more usually between about 40% andabout 60%, relative to the level present at day 7 after injection. Atday 40 after injection, the percentage in vivo vitreal level may begreater than about 20%, and more usually greater than about 40%,relative to the level present at day 7 after injection.

At day 45 after injection, the percentage in vivo vitreal level may bebetween about 15% and about 55%, and more usually between about 25% andabout 45%, relative to the level present at day 7 after injection. Atday 67 after injection, the percentage in vivo vitreal level may begreater than about 15%, and more usually greater than about 25%,relative to the level present at day 7 after injection.

At day 90 after injection, the percentage in vivo vitreal level may bebetween about 60% and about 100%, and more usually between about 70% andabout 90%, relative to the level present at day 7 after injection. Atday 90 after injection, the percentage in vivo vitreal level may begreater than about 60%, and more usually greater than about 70%,relative to the level present at day 7 after injection.

In some variations, the percentage in vivo vitreal level has thefollowing characteristics relative to the level present at day 7 afterinjection: at 32 days after injection it is less than about 80%; at 45days after injection it is less than about 60%; and 90 days afterinjection it is less than about 100%.

In some variations, the liquid formulation when injected between thesclera and conjunctiva of a rabbit eye delivers therapeutic agent givingan average concentration of therapeutic agent in the retina choroidtissues of the rabbit eye of at least about 0.1 μg/mg for at least about30, at least about 60, or at least about 90 days after administration ofthe liquid formulation to the rabbit eye. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the vitreous of the rabbit eye of at least about0.01 ng/mg for at least about 30, at least about 60, or at least about90 days after administration of the liquid formulation to the rabbiteye. In some variations, the liquid formulation when injected betweenthe sclera and conjunctiva of a rabbit eye delivers therapeutic agentgiving an average concentration of therapeutic agent in the vitreous ofthe rabbit eye of at least about 0.1 ng/mg for at least about 30, atleast about 60, or at least about 90 days after administration of theliquid formulation to the rabbit eye. In some variations, the liquidformulation when injected between the sclera and conjunctiva of a rabbiteye delivers therapeutic agent giving an average concentration oftherapeutic agent in the vitreous of the rabbit eye of at least about 1ng/mL for at least about 30, at least about 60, or at least about 90days after administration of the liquid formulation to the rabbit eye.

In some variations, the ratio of the base ten logarithms of the averagelevels of a therapeutic agent in two or more of the retina choroidtissues, the sclera, and the vitreous is approximately constant over anextended period of time after placement of the in situ gellingformulation in or proximate to the eye. In some variations, the ratio ofthe base ten logarithms of the average levels of a therapeutic agent intwo or more of the retina choroid tissues, the sclera, and the vitreousis approximately constant over an extended period of time afterplacement of the in situ gelling formulation between the sclera and theconjunctiva of an eye. In some variations, the ratio of the base tenlogarithms of the average levels of a therapeutic agent in the vitreousand the sclera is approximately constant over an extended period of timeafter placement of the in situ gelling formulation between the scleraand the conjunctiva of an eye.

In some variations, the ratio of the base ten logarithms of the averagelevels of a therapeutic agent in the vitreous and the retina choroidtissues is approximately constant over an extended period of time. Putanother way, as the level of therapeutic agent in the vitreous rises,the level of therapeutic agent in the retina choroid tissues rises to asimilar degree when considered on the logarithmic scale, and vice versa.

In some variations, the ratio of the base ten logarithms of the averagelevels of a therapeutic agent in the vitreous versus the retina choroidtissues is approximately constant over an extended period of time ofabout 7, about 30, about 60, or about 90 days. In some variations, theratio of the average level of therapeutic agent in the vitreous relativeto the level of therapeutic agent in the retina choroid tissues afterplacement of the in situ gelling formulation between the sclera and theconjunctiva of an eye is constant at about 37:1 at day 7, about 40:1 atday 32, about 10:1 at day 45, and about 34:1 at day 90.

In some variations, the ratio of the average level of therapeutic agentin the vitreous relative to the level of therapeutic agent in the retinachoroid tissues is constant at about 40:1 over a period of about 7,about 32, about 45, or about 90 days.

In some variations, the average level of the therapeutic agent in any orall of the retina choroid tissues, the sclera, and the vitreous isapproximately constant over an extended period of time after placementof the in situ gelling formulation in or proximate to the eye.

In some variations, after placement of an in situ gelling formulationbetween the sclera and the conjunctiva, the average level of therapeuticagent in the vitreous is approximately constant at about 8.1 ng/ml. Insome variations, after placement of an in situ gelling formulationbetween the sclera and the conjunctiva, the average level of therapeuticagent in the retina choroid tissues is approximately constant at about0.25 ng/mg. In some variations, after placement of an in situ gellingformulation between the sclera and the conjunctiva, the average level oftherapeutic agent in the sclera is approximately constant at about 1930ng/mg.

In some variations, the in situ gelling formulation when injectedbetween the sclera and conjunctiva of a rabbit eye maintains an averagelevel of therapeutic agent in the vitreous that is approximatelyconstant at about 0.1 μg/mL for at least about 30, at least about 60, orat least about 90 days after administration of the liquid formulation tothe rabbit eye. In some variations, the in situ gelling formulation wheninjected between the sclera and conjunctiva of a rabbit eye maintains anaverage level of therapeutic agent in the vitreous that is approximatelyconstant at about 0.001 ng/mL for at least about 30, at least about 60,or at least about 90 days after administration of the liquid formulationto the rabbit eye. In some variations, the in situ gelling formulationwhen injected between the sclera and conjunctiva of a rabbit eyemaintains an average level of therapeutic agent in the vitreous that isapproximately constant at about 0.01 ng/mL for at least about 30, atleast about 60, or at least about 90 days after administration of theliquid formulation to the rabbit eye. In some variations, the in situgelling formulation when injected between the sclera and conjunctiva ofa rabbit eye maintains an average level of therapeutic agent in thevitreous that is approximately constant at about 0.1 ng/mL for at leastabout 30, at least about 60, or at least about 90 days afteradministration of the liquid formulation to the rabbit eye. In somevariations, the in situ gelling formulation when injected between thesclera and conjunctiva of a rabbit eye maintains an average level oftherapeutic agent in the vitreous that is approximately constant atabout 1 ng/mL for at least about 30, at least about 60, or at leastabout 90 days after administration of the liquid formulation to therabbit eye. In some variations, the in situ gelling formulation wheninjected between the sclera and conjunctiva of a rabbit eye maintains anaverage level of therapeutic agent in the vitreous that is approximatelyconstant at about 10 ng/mL for at least about 30, at least about 60, orat least about 90 days after administration of the liquid formulation tothe rabbit eye. In some variations, the in situ gelling formulation wheninjected between the sclera and conjunctiva of a rabbit eye maintains anaverage level of therapeutic agent in the vitreous that is approximatelyconstant at about 100 ng/mL for at least about 30, at least about 60, orat least about 90 days after administration of the liquid formulation tothe rabbit eye.

In some variations, the in situ gelling formulation when injectedbetween the sclera and conjunctiva of a rabbit eye maintains an averagelevel of therapeutic agent in the retina choroid tissues that isapproximately constant at about 0.1 μg/mg for at least about 30, atleast about 60, or at least about 90 days after administration of theliquid formulation to the rabbit eye. In some variations, the in situgelling formulation when injected between the sclera and conjunctiva ofa rabbit eye maintains an average level of therapeutic agent in theretina choroid tissues that is approximately constant at about 0.001ng/mg for at least about 30, at least about 60, or at least about 90days after administration of the liquid formulation to the rabbit eye.In some variations, the in situ gelling formulation when injectedbetween the sclera and conjunctiva of a rabbit eye maintains an averagelevel of therapeutic agent in the retina choroid tissues that isapproximately constant at about 0.01 ng/mg for at least about 30, atleast about 60, or at least about 90 days after administration of theliquid formulation to the rabbit eye. In some variations, the in situgelling formulation when injected between the sclera and conjunctiva ofa rabbit eye maintains an average level of therapeutic agent in theretina choroid tissues that is approximately constant at about 0.1 ng/mgfor at least about 30, at least about 60, or at least about 90 daysafter administration of the liquid formulation to the rabbit eye. Insome variations, the in situ gelling formulation when injected betweenthe sclera and conjunctiva of a rabbit eye maintains an average level oftherapeutic agent in the retina choroid tissues that is approximatelyconstant at about 1 ng/mg for at least about 30, at least about 60, orat least about 90 days after administration of the liquid formulation tothe rabbit eye. In some variations, the in situ gelling formulation wheninjected between the sclera and conjunctiva of a rabbit eye maintains anaverage level of therapeutic agent in the retina choroid tissues that isapproximately constant at about 10 ng/mg for at least about 30, at leastabout 60, or at least about 90 days after administration of the liquidformulation to the rabbit eye.

In some variations, the in situ gelling formulation when injectedbetween the sclera and conjunctiva of a rabbit eye maintains an averagelevel of therapeutic agent in the sclera that is approximately constantat about 0.1 μg/mg for at least about 30, at least about 60, or at leastabout 90 days after administration of the liquid formulation to therabbit eye. In some variations, the in situ gelling formulation wheninjected between the sclera and conjunctiva of a rabbit eye maintains anaverage level of therapeutic agent in the sclera that is approximatelyconstant at about 0.001 ng/mg for at least about 30, at least about 60,or at least about 90 days after administration of the liquid formulationto the rabbit eye. In some variations, the in situ gelling formulationwhen injected between the sclera and conjunctiva of a rabbit eyemaintains an average level of therapeutic agent in the sclera that isapproximately constant at about 0.01 ng/mg for at least about 30, atleast about 60, or at least about 90 days after administration of theliquid formulation to the rabbit eye. In some variations, the in situgelling formulation when injected between the sclera and conjunctiva ofa rabbit eye maintains an average level of therapeutic agent in thesclera that is approximately constant at about 0.1 ng/mg for at leastabout 30, at least about 60, or at least about 90 days afteradministration of the liquid formulation to the rabbit eye. In somevariations, the in situ gelling formulation when injected between thesclera and conjunctiva of a rabbit eye maintains an average level oftherapeutic agent in the sclera that is approximately constant at about1 ng/mg for at least about 30, at least about 60, or at least about 90days after administration of the liquid formulation to the rabbit eye.In some variations, the in situ gelling formulation when injectedbetween the sclera and conjunctiva of a rabbit eye maintains an averagelevel of therapeutic agent in the sclera that is approximately constantat about 10 ng/mg for at least about 30, at least about 60, or at leastabout 90 days after administration of the liquid formulation to therabbit eye. In some variations, the in situ gelling formulation wheninjected between the sclera and conjunctiva of a rabbit eye maintains anaverage level of therapeutic agent in the sclera that is approximatelyconstant at about 100 ng/mg for at least about 30, at least about 60, orat least about 90 days after administration of the liquid formulation tothe rabbit eye. In some variations, the in situ gelling formulation wheninjected between the sclera and conjunctiva of a rabbit eye maintains anaverage level of therapeutic agent in the sclera that is approximatelyconstant at about 1 μg/mg for at least about 30, at least about 60, orat least about 90 days after administration of the liquid formulation tothe rabbit eye. In some variations, the in situ gelling formulation wheninjected between the sclera and conjunctiva of a rabbit eye maintains anaverage level of therapeutic agent in the sclera that is approximatelyconstant at about 10 μg/mg for at least about 30, at least about 60, orat least about 90 days after administration of the liquid formulation tothe rabbit eye.

For treatment, prevention, inhibition, delaying the onset of, or causingthe regression of certain diseases or conditions, it may be desirable tomaintain delivery of a therapeutically effective amount of thetherapeutic agent for an extended period of time. Depending on thedisease or condition being treated, prevented, inhibited, having onsetdelayed, or being caused to regress this extended period of time may beat least about 1 week, at least about 2 weeks, at least about 3 weeks,at least about 1 month, at least about 3 months, at least about 6months, at least about 9 months, or at least about 1 year. Generally,however, any extended period of delivery may be possible. Atherapeutically effective amount of agent may be delivered for anextended period by a liquid formulation or composition that maintainsfor the extended period a concentration of agent in a subject or an eyeof a subject sufficient to deliver a therapeutically effective amount ofagent for the extended time.

Delivery of a therapeutically effective amount of the therapeutic agentfor an extended period may be achieved via placement of one compositionor liquid formulation or may be achieved by application of two or moredoses of composition or liquid formulations. As a non-limiting exampleof such multiple applications, maintenance of the therapeutic amount ofrapamycin for 3 months for treatment, prevention, inhibition, delay ofonset, or cause of regression of wet AMD may be achieved by applicationof one liquid formulation or composition delivering a therapeutic amountfor 3 months or by sequential application of a plurality of liquidformulations or compositions. The optimal dosage regime will depend onthe therapeutic amount of the therapeutic agent needing to be delivered,and the period over which it need be delivered. Those versed in suchextended therapeutic agent delivery dosing will understand how toidentify dosing regimes that may be used based on the teachings providedherein.

When using certain therapeutic agents or for the treatment, prevention,inhibition, delaying the onset of, or causing the regression of certaindiseases, it may be desirable for delivery of the therapeutic agent notto commence immediately upon placement of the liquid formulation orcomposition into the eye region, but for delivery to commence after somedelay. For example, but in no way limiting, such delayed release may beuseful where the therapeutic agent inhibits or delays wound healing anddelayed release is desirable to allow healing of any wounds occurringupon placement of the liquid formulation or composition. Depending onthe therapeutic agent being delivered and/or the diseases and conditionsbeing treated, prevented, inhibited, onset delayed, and regressioncaused this period of delay before delivery of the therapeutic agentcommences may be about 1 hour, about 6 hours, about 12 hours, about 18hours, about 1 day, about 2 days, about 3 days, about 4 days, about 5days, about 6 days, about 7 days, about 8 days, about 9 days, about 10days, about 11 days, about 12 days, about 13 days, about 14 days, about21 days, about 28 days, about 35 days, or about 42 days. Other delayperiods may be possible. Delayed release formulations that may be usedare known to people versed in the technology.

Intravitreal, Subconjunctival, Subtenonal, and Topical Delivery ofRapamycin for Treatment, Prevention, Inhibition, Delay of Onset, orCause of Regression of AMD

In one method described herein, a liquid formulation comprisingrapamycin is delivered subconjunctivally or to the vitreous of an eye toprevent, treat, inhibit, delay onset of, or cause regression ofangiogenesis in the eye, including but not limited to treating CNV asobserved, for example, in AMD. In some variations, the liquidformulation is used to treat angiogenesis in the eye, including but notlimited to treating CNV as observed, for example, in AMD. Rapamycin hasbeen shown to inhibit CNV in rat and mice models, as described in U.S.application Ser. No. 10/665,203, which is incorporated herein byreference in its entirety. Rapamycin has been observed to inhibitMatrigel™ and laser-induced CNV when administered systemically andsubretinally. Also, periocular injection of rapamycin inhibitslaser-induced CNV.

Other therapeutic agents that may be delivered to the eye, particularlythe vitreous of an eye, for treatment, prevention, inhibition, delayingonset, or causing regression of angiogenesis in the eye (such as CNV)are members of the limus family of compounds other than rapamycinincluding but not limited to everolimus and tacrolimus (FK-506).

As described herein, the dosage of the therapeutic agent will depend onthe condition being addressed, whether the condition is to be treated,prevented, inhibited, have onset delayed, or be caused to regress, theparticular therapeutic agent, and other clinical factors such as weightand condition of the subject and the route of administration of thetherapeutic agent. It is to be understood that the methods, liquidformulations, and compositions described herein have application forboth human and veterinary use, as well as uses in other possibleanimals. As described herein, tissue concentrations of therapeuticagents expressed in units of mass per volume generally refer to tissuesthat are primarily aqueous such as the vitreous, for example. Tissueconcentrations of therapeutic agents expressed in unit of mass per massgenerally refer to other tissues such as the sclera or retina choroidtissues, for example.

One concentration of rapamycin that may be used in the methods describedherein is one that provides about 0.01 pg/ml or pg/mg or more ofrapamycin at the tissue level. Another concentration that may be used isone that provides about 0.1 pg/ml or ng/mg or more at the tissue level.Another concentration that may be used is one that provides about 1pg/ml or ng/mg or more at the tissue level. Another concentration thatmay be used is one that provides about 0.01 ng/ml or ng/mg or more atthe tissue level. Another concentration that may be used is one thatprovides about 0.1 ng/ml or ng/mg or more at the tissue level. Anotherconcentration that may be used is one that provides about 0.5 ng/ml orng/mg or more at the tissue level. Another concentration that may beused is one that provides about 1 ng/ml or more at the tissue level.Another concentration that may be used is one that provides about 2ng/ml or more at the tissue level. Another concentration that may beused is one that provides about 3 ng/ml or more at the tissue level.Another concentration that may be used is one that provides about 5ng/ml or more at the tissue level. Another concentration that may beused is one that provides about 10 ng/ml or more at the tissue level.Another concentration that may be used is one that provides about 15ng/ml or more at the tissue level. Another concentration that may beused is one that provides about 20 ng/ml or more at the tissue level.Another concentration that may be used is one that provides about 30ng/ml or more at the tissue level. Another concentration that may beused is one that provides about 50 ng/ml or more at the tissue level.One of ordinary skill in the art would know how to arrive at anappropriate concentration depending on the route and duration ofadministration utilized, given the teachings herein.

Generally, the amount of rapamycin administered in a liquid formulationis an amount sufficient to treat, prevent, inhibit, delay the onset, orcause regression of the disease or condition of the eye for the requiredamount of time. In some variations the amount of rapamycin administeredin the liquid formulation is an amount sufficient to treat the diseaseor condition of the eye for the required amount of time.

In some variations, a total amount of rapamycin less than about 5 mg isadministered subconjunctivally. In some variations, a total amount ofrapamycin less than about 5.0 mg is administered subconjunctivally. Insome variations, a total amount of rapamycin less than about 4.5 mg isadministered subconjunctivally. In some variations, a total amount ofrapamycin less than about 4.0 mg is administered subconjunctivally. Insome variations, a total amount of rapamycin less than about 3.5 mg isadministered subconjunctivally. In some variations, a total amount ofrapamycin less than about 3.0 mg is administered subconjunctivally. Insome variations, a total amount of rapamycin less than about 2.5 mg isadministered subconjunctivally. In some variations, a total amount ofrapamycin less than about 2 mg is administered subconjunctivally. Insome variations, a total amount of rapamycin less than about 1.2 mg isadministered subconjunctivally. In some variations, a total amount ofrapamycin less than about 1.0 mg is administered subconjunctivally. Insome variations, a total amount of rapamycin less than about 0.8 mg isadministered subconjunctivally. In some variations, a total amount ofrapamycin less than about 0.6 mg is administered subconjunctivally. Insome variations, a total amount of rapamycin less than about 0.4 mg isadministered subconjunctivally. In some variations, a volume of aformulation is administered that contains an amount of rapamycindescribed herein.

In some variations, a liquid formulation containing a concentration ofrapamycin by weight of the total of between about 0.5% and about 6% issubconjunctivally administered to a human subject by administeringbetween about 0.1 μl and about 200 μl of a liquid formulation describedherein. In some variations, a liquid formulation containing aconcentration of rapamycin by weight of the total of between about 0.5%and about 4% is subconjunctivally administered to a human subject byadministering between about 1 μl and about 50 μl of a liquid formulationdescribed herein. In some variations, a liquid formulation containing aconcentration of rapamycin by weight of the total of between about 1.5%and about 3.5% is subconjunctivally administered to a human subject byadministering between about 1 μl and about 15 μl of a liquid formulationdescribed herein. In some variations, a liquid formulation containing aconcentration of rapamycin by weight of the total of about 2% issubconjunctivally administered to a human subject by administeringbetween about 1 μl and about 15 μl of a liquid formulation describedherein.

In some variations, a liquid formulation containing an amount ofrapamycin of between about 0.2 μg and about 4 mg is subconjunctivallyadministered to a human subject by administering between about 0.1 μland about 200 μl of a liquid formulation described herein. In somevariations, a liquid formulation containing an amount of rapamycin ofbetween about 20 μg and about 2 mg is subconjunctivally administered toa human subject by administering between about 1 μl and about 100 μl ofa liquid formulation described herein. In some variations, a liquidformulation containing an amount of rapamycin of between about 20 μg andabout 1 mg is subconjunctivally administered to a human subject byadministering between about 1 μl and about 50 μl of a liquid formulationdescribed herein. In some variations, a liquid formulation containing anamount of rapamycin of between about 20 μg and about 500 μg issubconjunctivally administered to a human subject by administeringbetween about 1 μl and about 25 μl of a liquid formulation describedherein. In some variations, a liquid formulation containing an amount ofrapamycin of between about 20 μg and about 300 μg is subconjunctivallyadministered to a human subject by administering between about 1 μl andabout 15 μl of a liquid formulation described herein.

In some variations, a total amount of rapamycin less than about 200 μgis administered intravitreally. In some variations, a total amount ofrapamycin less than about 200 μg is administered intravitreally. In somevariations, a total amount of rapamycin less than about 300 μg isadministered intravitreally. In some variations, a total amount ofrapamycin less than about 400 μg is administered intravitreally. In somevariations, a total amount of rapamycin less than about 500 μg isadministered intravitreally. In some variations, a total amount ofrapamycin less than about 600 μg is administered intravitreally. In somevariations, a total amount of rapamycin less than about 800 μg isadministered intravitreally. In some variations, a total amount ofrapamycin less than about 1 mg is administered intravitreally. In somevariations, a total amount of rapamycin less than about 2 mg isadministered intravitreally. In some variations, a total amount ofrapamycin less than about 2.5 mg is administered intravitreally. In somevariations, a total amount of rapamycin less than about 3 mg isadministered intravitreally. In some variations, a total amount ofrapamycin less than about 3.5 mg is administered intravitreally. In somevariations, a total amount of rapamycin less than about 4 mg isadministered intravitreally. In some variations, a volume of aformulation is administered that contains an amount of rapamycindescribed herein.

In some variations, a liquid formulation containing a concentration ofrapamycin by weight of the total of between about 0.5% and about 6% isintravitreally administered to a human subject by administering betweenabout 0.1 μl and about 200 μl of a liquid formulation described herein.In some variations, a liquid formulation containing a concentration ofrapamycin by weight of the total of between about 0.5% and about 4% isintravitreally administered to a human subject by administering betweenabout 1 μl and about 50 μl of a liquid formulation described herein. Insome variations, a liquid formulation containing a concentration ofrapamycin by weight of the total of between about 1.5% and about 3.5% isintravitreally administered to a human subject by administering betweenabout 1 μl and about 15 μl of a liquid formulation described herein. Insome variations, a liquid formulation containing a concentration ofrapamycin by weight of the total of about 2% is intravitreallyadministered to a human subject by administering between about 1 μl andabout 15 μl of a liquid formulation described herein.

In some variations, a liquid formulation containing an amount ofrapamycin of between about 0.2 μg and about 4 mg is intravitreallyadministered to a human subject by administering between about 0.1 μland about 200 μl of a liquid formulation described herein. In somevariations, a liquid formulation containing an amount of rapamycin ofbetween about 20 μg and about 2 mg is intravitreally administered to ahuman subject by administering between about 1 μl and about 100 μl of aliquid formulation described herein. In some variations, a liquidformulation containing an amount of rapamycin of between about 20 μg andabout 1 mg is intravitreally administered to a human subject byadministering between about 1 μl and about 50 μl of a liquid formulationdescribed herein. In some variations, a liquid formulation containing anamount of rapamycin of between about 20 μg and about 500 μg isintravitreally administered to a human subject by administering betweenabout 1 μl and about 25 μl of a liquid formulation described herein. Insome variations, a liquid formulation containing an amount of rapamycinof between about 20 μg and about 300 μg is intravitreally administeredto a human subject by administering between about 1 μl and about 15 μlof a liquid formulation described herein.

In some variations, a total amount of rapamycin less than about 5 mg isadministered subtenonally. In some variations, a total amount ofrapamycin less than about 5.0 mg is administered subtenonally. In somevariations, a total amount of rapamycin less than about 4.5 mg isadministered subtenonally. In some variations, a total amount ofrapamycin less than about 4.0 mg is administered subtenonally. In somevariations, a total amount of rapamycin less than about 3.5 mg isadministered subtenonally. In some variations, a total amount ofrapamycin less than about 3.0 mg is administered subtenonally. In somevariations, a total amount of rapamycin less than about 2.5 mg isadministered subtenonally. In some variations, a total amount ofrapamycin less than about 2 mg is administered subtenonally. In somevariations, a total amount of rapamycin less than about 1.2 mg isadministered subtenonally. In some variations, a total amount ofrapamycin less than about 1.0 mg is administered subtenonally. In somevariations, a total amount of rapamycin less than about 0.8 mg isadministered subtenonally. In some variations, a total amount ofrapamycin less than about 0.6 mg is administered subtenonally. In somevariations, a total amount of rapamycin less than about 0.4 mg isadministered subtenonally. In some variations, a volume of a formulationis administered that contains an amount of therapeutic agent describedherein.

In some variations, a total amount of rapamycin less than about 5 mg isadministered topically. In some variations, a total amount of rapamycinless than about 5.0 mg is administered topically. In some variations, atotal amount of rapamycin less than about 4.5 mg is administeredtopically. In some variations, a total amount of rapamycin less thanabout 4.0 mg is administered topically. In some variations, a totalamount of rapamycin less than about 3.5 mg is administered topically. Insome variations, a total amount of rapamycin less than about 3.0 mg isadministered topically. In some variations, a total amount of rapamycinless than about 2.5 mg is administered topically. In some variations, atotal amount of rapamycin less than about 2 mg is administeredtopically. In some variations, a total amount of rapamycin less thanabout 1.2 mg is administered topically. In some variations, a totalamount of rapamycin less than about 1.0 mg is administered topically. Insome variations, a total amount of rapamycin less than about 0.8 mg isadministered topically. In some variations, a total amount of rapamycinless than about 0.6 mg is administered topically. In some variations, atotal amount of rapamycin less than about 0.4 mg is administeredtopically. In some variations, a volume of a formulation is administeredthat contains an amount of rapamycin described herein. In somevariations, a total amount of therapeutic agent administered topicallyis any of between about 20 μg and about 4000 μg, between about 10 μg andabout 2000 μg, between about 10 μg and 1750 μg, between about 1500 μgand 1000 μg, or between about 10 μg and 1000 μg. In some variations, atotal amount of therapeutic agent administered topically is about 1660μg. In some variations, a total amount of therapeutic agent administeredtopically is about 880 μg. In some variations, a total amount oftherapeutic agent administered topically is 40 μg. In some variations, atotal amount of therapeutic agent administered topically is about 28 μg.

In some variations, the liquid formulation when topically administeredto a rabbit eye delivers therapeutic agent giving an averageconcentration of therapeutic agent in the cornea of the rabbit eye of atleast any of about 0.001 ng/mg, 0.01 ng/mg, 0.1 ng/mg, or 1 ng/mg. Insome variations, the liquid formulation when topically administered to arabbit eye delivers therapeutic agent giving an average concentration oftherapeutic agent in the cornea of the rabbit eye of at least 0.01ng/mg. In some variations, the liquid formulation when topicallyadministered to a rabbit eye delivers therapeutic agent giving anaverage concentration of therapeutic agent in the retina choroid tissuesof the rabbit eye of at least any of about 0.0001 ng/mg, 0.001 ng/mg,0.01 ng/mg, 0.1 ng/mg, or 1 ng/mg. In some variations, the liquidformulation when topically administered to a rabbit eye deliverstherapeutic agent giving an average concentration of therapeutic agentin the retina choroid tissues of the rabbit eye of at least 0.001 ng/mg.

In some variations, a total amount of rapamycin administeredsubconjunctivally is any of between about 50 μg and about 3 mg, betweenabout 150 μg and about 750 μg, between about 300 μg and about 1000 μg,between about 300 μg and about 950 μg, between about 400 μg and about900 μg, between about 450 μg and about 850 μg, between about 500 μg andabout 800 μg, between about 550 μg and about 750 μg, or between about600 μg and about 700 μg. In some variations, a total amount of rapamycinadministered subconjunctivally is any of about 220 μg, about 440 μg,about 587 μg, about 630 μg, about 660 μg, about 880 μg, about 1320 μg,about 1760 μg, or about 2200 μg. In some variations, a total amount ofrapamycin administered subconjunctivally is about 220 μg. In somevariations, a total amount of rapamycin administered subconjunctivallyis about 440 μg. In some variations, a total amount of rapamycinadministered subconjunctivally is about 660 μg. In some variations, atotal amount of rapamycin administered subconjunctivally is about 880μg. In some variations, a liquid formulation containing an amount ofrapamycin of 220 μg is subconjunctivally administered to a human subjectby administering about 10 μl of a liquid formulation described herein.In some variations, a liquid formulation containing an amount ofrapamycin of 440 μg is subconjunctivally administered to a human subjectby administering about 20 μl of a liquid formulation described herein.In some variations, a liquid formulation containing an amount ofrapamycin of 660 μg is subconjunctivally administered to a human subjectby administering about 30 μl of a liquid formulation described herein.In some variations, a liquid formulation containing an amount ofrapamycin of 880 μg is subconjunctivally administered to a human subjectby administering about 40 μl of a liquid formulation described herein.

In some variations, a total amount of rapamycin administeredintravitreally is any of between about 20 μg and about 750 μg, betweenabout 20 μg and about 500 μg, or between about 30 μg and about 200 μg.In some variations, a total amount of rapamycin administeredintravitreally is any of about 44 μg, about 110 μg, about 132 μg, about133.5 μg, about 176 μg, about 264 μg, about 352 μg, or about 440 μg. Insome variations, a total amount of rapamycin administered intravitreallyis about 44 μg. In some variations, a total amount of rapamycinadministered intravitreally is about 110 μg. In some variations, aliquid formulation containing an amount of rapamycin of 44 μg isintravitreally administered to a human subject by administering about 2μl of a liquid formulation described herein. In some variations, aliquid formulation containing an amount of rapamycin of 110 μg isintravitreally administered to a human subject by administering about 5μl of a liquid formulation described herein. In some variations, aliquid formulation containing an amount of rapamycin of 176 μg isintravitreally administered to a human subject by administering about 8μl of a liquid formulation described herein.

In some variations a liquid formulation as described herein containingan amount of rapamycin of between about 1 μg and about 5 mg isadministered to a human subject for treatment of wet AMD. In somevariations a liquid formulation as described herein containing an amountof rapamycin of between about 20 μg and about 4 mg is administered to ahuman subject for treatment of wet AMD. In some variations a liquidformulation as described herein containing an amount of rapamycin ofbetween about 20 μg and about 1.2 mg is administered to a human subjectfor treatment of wet AMD. In some variations an amount of rapamycin ofbetween about 10 μg and about 0.5 mg is administered to a human subjectfor treatment of wet AMD. In some variations an amount of rapamycin ofbetween about 10 μg and 90 μg is administered to a human subject fortreatment of wet AMD. In some variations an amount of rapamycin ofbetween about 60 μg and about 120 μg is administered to a human subjectfor treatment of wet AMD. In some variations an amount of rapamycin ofbetween about 100 μg and about 400 μg is administered to a human subjectfor treatment of wet AMD. In some variations an amount of rapamycin ofbetween about 400 μg and about 1 mg is administered to a human subjectfor treatment of wet AMD. In some variations an amount of rapamycin ofbetween about 1 mg and about 5 mg is administered to a human subject fortreatment of wet AMD. In some variations, an amount of rapamycin ofbetween about 3 mg and about 7 mg is administered to a human subject fortreatment of wet AMD. In some variations, an amount of rapamycin ofbetween about 5 mg and about 10 mg is administered to a human subjectfor treatment of wet AMD.

In some variations a liquid formulation as described herein containingan amount of rapamycin of between about 1 μg and about 5 mg isadministered to a human subject for prevention of wet AMD. In somevariations a liquid formulation as described herein containing an amountof rapamycin of between about 20 μg and about 4 mg is administered to ahuman subject for prevention of wet AMD. In some variations a liquidformulation as described herein containing an amount of rapamycin ofbetween about 20 μg and about 1.2 mg is administered to a human subjectfor prevention of wet AMD. In some variations an amount of rapamycin ofbetween about 10 μg and about 0.5 mg is administered to a human subjectfor prevention of wet AMD. In some variations an amount of rapamycin ofbetween about 10 μg and 90 μg is administered to a human subject forprevention of wet AMD. In some variations an amount of rapamycin ofbetween about 60 μg and about 120 μg is administered to a human subjectfor prevention of wet AMD. In some variations an amount of rapamycin ofbetween about 100 μg and about 400 μg is administered to a human subjectfor prevention of wet AMD. In some variations an amount of rapamycin ofbetween about 400 μg and about 1 mg is administered to a human subjectfor prevention of wet AMD. In some variations an amount of rapamycin ofbetween about 1 mg and about 5 mg is administered to a human subject forprevention of wet AMD. In some variations, an amount of rapamycin ofbetween about 3 mg and about 7 mg is administered to a human subject forprevention of wet AMD. In some variations, an amount of rapamycin ofbetween about 5 mg and about 10 mg is administered to a human subjectfor prevention of wet AMD.

In some variations a liquid formulation as described herein containingan amount of rapamycin of between about 1 μg and about 5 mg isadministered to a human subject for treatment of dry AMD. In somevariations a liquid formulation as described herein containing an amountof rapamycin of between about 20 μg and about 4 mg is administered to ahuman subject for treatment of dry AMD. In some variations a liquidformulation as described herein containing an amount of rapamycin ofbetween about 20 μg and about 1.2 mg is administered to a human subjectfor treatment of dry AMD. In some variations an amount of rapamycin ofbetween about 10 μg and about 0.5 mg is administered to a human subjectfor treatment of dry AMD. In some variations an amount of rapamycin ofbetween about 10 μg and 90 μg is administered to a human subject fortreatment of dry AMD. In some variations an amount of rapamycin ofbetween about 60 μg and about 120 μg is administered to a human subjectfor treatment of dry AMD. In some variations an amount of rapamycin ofbetween about 100 μg and about 400 μg is administered to a human subjectfor treatment of dry AMD. In some variations an amount of rapamycin ofbetween about 400 μg and about 1 mg is administered to a human subjectfor treatment of dry AMD. In some variations an amount of rapamycin ofbetween about 1 mg and about 5 mg is administered to a human subject fortreatment of dry AMD. In some variations, an amount of rapamycin ofbetween about 3 mg and about 7 mg is administered to a human subject fortreatment of dry AMD. In some variations, an amount of rapamycin ofbetween about 5 mg and about 10 mg is administered to a human subjectfor treatment of dry AMD.

In some variations, a liquid formulation as described herein containingan amount of rapamycin of between about 1 μg and about 5 mg isadministered to a human subject for treatment of angiogenesis, includingbut not limited to choroidal neovascularization. In some variations, anamount of rapamycin of between about 20 μg and about 4 mg isadministered to the human subject; between about 20 μg and about 1.2 mg;between about 10 μg and about 0.5 mg is administered to a human subjectfor treatment of wet AMD, between about 10 μg and 90 μg, between about60 μg and 120 μg is administered to the human subject; between about 100μg and 400 μg, between about 400 μg and 1 mg is administered to thehuman subject; in some variations, an amount of rapamycin of betweenabout 1 mg and 5 mg is administered to the human subject; in somevariations, an amount of rapamycin of between about 3 mg and 7 mg isadministered to the human subject; in some variations, an amount ofrapamycin of between about 5 mg and 10 mg is administered to the humansubject for treatment of angiogenesis, including but not limited tochoroidal neovascularization.

In one method, a liquid formulation as described herein contains anamount of a therapeutic agent equivalent to an amount of rapamycin.

In one method, a liquid formulation as described herein containing anamount of a therapeutic agent equivalent to an amount of rapamycin ofbetween about 1 μg and about 5 mg is administered to a human subject fortreatment of wet AMD. In some variations, an amount of a therapeuticagent equivalent to an amount of rapamycin of between about 1 μg andabout 5 mg is administered to the human subject; between about 20 μg andabout 1.2 mg; between about 10 μg and about 0.5 mg is administered to ahuman subject for treatment of wet AMD, between about 10 μg and 90 μg,between about 60 μg and 120 μg is administered to the human subject;between about 100 μg and 400 μg, between about 400 μg and 1 mg isadministered to the human subject is administered to the human subject;in some variations, an amount of a therapeutic agent equivalent to anamount of rapamycin of between about 1 mg and 5 mg is administered tothe human subject; in some variations, an amount of a therapeutic agentequivalent to an amount of rapamycin of between about 3 mg and 7 mg isadministered to the human subject; in some variations, an amount of atherapeutic agent equivalent to an amount of rapamycin of between about5 mg and 10 mg is administered to the human subject.

In some variations, a liquid formulation as described herein containingan amount of a therapeutic agent equivalent to an amount of rapamycin ofbetween about 1 μg and about 5 mg is administered to a human subject fortreatment of dry AMD. In some variations, an amount of a therapeuticagent equivalent to an amount of rapamycin of between about 20 μg andabout 4 mg is administered to the human subject; between about 20 μg andabout 1.2 mg; between about 10 μg and about 0.5 mg is administered to ahuman subject for treatment of wet AMD, between about 10 μg and 90 μg,between about 60 μg and 120 μg is administered to the human subject;between about 100 μg and 400 μg, between about 400 μg and 1 mg isadministered to the human subject; in some variations, an amount of atherapeutic agent equivalent to an amount of rapamycin of between about400 μg and 1 mg is administered to the human subject; in somevariations, an amount of a therapeutic agent equivalent to an amount ofrapamycin of between about 1 mg and 5 mg is administered to the humansubject; in some variations, an amount of a therapeutic agent equivalentto an amount of rapamycin of between about 3 mg and 7 mg is administeredto the human subject; in some variations, an amount of a therapeuticagent equivalent to an amount of rapamycin of between about 5 mg and 10mg is administered to the human subject to treat dry AMD.

In some variations, a liquid formulation as described herein containingan amount of a therapeutic agent equivalent to an amount of rapamycin ofbetween about 1 μg and about 5 mg is administered to a human subject forprevention of wet AMD. In some variations, an amount of a therapeuticagent equivalent to an amount of rapamycin of between about 20 μg andabout 4 mg is administered to the human subject; between about 20 μg andabout 1.2 mg; between about 10 μg and about 0.5 mg is administered to ahuman subject for prevention of wet AMD, between about 10 μg and 90 μg,between about 60 μg and 120 μg is administered to the human subject;between about 100 μg and 400 μg, between about 400 μg and 1 mg isadministered to the human subject; in some variations, an amount of atherapeutic agent equivalent to an amount of rapamycin of between about400 μg and 1 mg is administered to the human subject; in somevariations, an amount of a therapeutic agent equivalent to an amount ofrapamycin of between about 1 mg and 5 mg is administered to the humansubject; in some variations, an amount of a therapeutic agent equivalentto an amount of rapamycin of between about 3 mg and 7 mg is administeredto the human subject; in some variations, an amount of a therapeuticagent equivalent to an amount of rapamycin of between about 5 mg and 10mg is administered to the human subject to prevent wet AMD.

In some variations, any one or more of the formulations described hereinare administered intravitreally every 3 or more months, every 6 or moremonths, every 9 or more months, or every 12 or more months, or longer,to treat one or more of choroidal neovascularization, wet AMD, dry AMD,to prevent wet AMD, or to prevent progression of dry AMD to wet AMD. Insome variations, any one or more of the formulations described hereinare administered subconjunctivally every 3 or more months, every 6 ormore months, every 9 or more months, or every 12 or more months, orlonger, to treat one or more of choroidal neovascularization, wet AMD,dry AMD, or to prevent wet AMD.

In some variations, any one or more of the rapamycin formulationsdescribed herein are administered intravitreally every 3 or more months,every 6 or more months, every 9 or more months, or every 12 or moremonths, or longer, to treat one or more of choroidal neovascularization,wet AMD, dry AMD, to prevent wet AMD, or to prevent progression of dryAMD to wet AMD. In some variations, any one or more of the rapamycinformulations described herein are administered subconjunctivally every 3or more months, every 6 or more months, every 9 or more months, or every12 or more months, or longer, to treat one or more of choroidalneovascularization, wet AMD, dry AMD, or to prevent wet AMD. In somevariations, the effect of the rapamycin persists beyond the periodduring which it is present in the ocular tissues.

Delivery of the therapeutic agents described herein may, for example, bedelivered at a dosage range between about 1 ng/day and about 100 μg/day,or at dosages higher or lower than this range, depending on the routeand duration of administration. In some variations of liquid formulationor composition used in the methods described herein, the therapeuticagents are delivered at a dosage range of between about 0.1 μg/day andabout 10 μg/day. In some variations of liquid formulation or compositionused in the methods described herein, the therapeutic agents aredelivered at a dosage range of between about 1 μg/day and about 5μg/day. Dosages of various therapeutic agents for treatment, prevention,inhibition, delay of onset, or cause of regression of various diseasesand conditions described herein can be refined by the use of clinicaltrials.

The liquid formulations, including but not limited to solutions,suspensions, emulsions and situ gelling formulations, and compositionsdescribed herein may be used for delivery to the eye, as one nonlimitingexample by ocular or periocular administration, of therapeuticallyeffective amounts of rapamycin for extended periods of time to treat,prevent, inhibit, delay the onset of, or cause regression of CNV, andthus may be used to treat, prevent, inhibit, delay the onset of, orcause regression of wet AMD, or transition of dry AMD to wet AMD. It isbelieved that by changing certain characteristics of the liquidformulations described herein, including but not limited to thecomponents of the liquid formulations, the location in the eye to whichthe liquid formulation is delivered, including without limitationsubconjunctival or intravitreal placement, the liquid formulations maybe used to deliver therapeutically effective amounts of rapamycin to theeye for a variety of extended time periods including delivery oftherapeutic amounts for greater than about 1 week, for greater thanabout 2 weeks, for greater than about 3 weeks, for greater than about 1month, for greater than about 3 months, for greater than about 6 months,for greater than about 9 months, for greater than about 1 year.

When a therapeutically effective amount of rapamycin is administered toa subject suffering from wet AMD, the rapamycin may treat, inhibit, orcause regression of the wet AMD. Different therapeutically effectiveamounts may be required for treatment, inhibition or causing regression.A subject suffering from wet AMD may have CNV lesions, and it isbelieved that administration of a therapeutically effective amount ofrapamycin may have a variety of effects, including but not limited tocausing regression of the CNV lesions, stabilizing the CNV lesion, andpreventing progression of an active CNV lesion.

When a therapeutically effective amount of rapamycin is administered toa subject suffering from dry AMD, it is believed that the rapamycin mayprevent or slow the progression of dry AMD to wet AMD.

EXAMPLES

Unless the context indicates otherwise, the error bars in the chartsshow one standard deviation. Where ethanol is used, it is 200 proofethanol from Gold Shield Distributors, Hayward, Calif. Where rapamycinis used, it is from LC laboratories, Woburn, Mass., or Chunghwa ChemicalSynthesis & Biotech Co., LTD (CCSB), Taipei Hsien, Taiwan, ROC. WherePEG 400 is used, it is from The Dow Chemical Company, New Milford, Conn.Some of the graphs use the expression “uL” or “ug” to refer to μL or μg,respectively. Where a volume of 10 μL or less is administered, HamiltonHPLC syringes were used.

Examples 1-45 Preparation and Administration of Rapamycin-ContainingCompositions

Preparation and characterization of exemplary rapamycin-containingcompositions, and administration thereof to New Zealand white rabbits isdescribed in US 2006/0258698 A1 and WO 2006/102378A2 of Mudumba et al.(Examples 1-45 and associated figures herein incorporated by reference).

Example 46 Preparation and Characterization of a Rapamycin-ContainingSolution

About 320 g of ethanol was sparged with N₂ for about 10 minutes, andthen about 40 g of sirolimus was added to the ethanol. The mixture wassonicated for about 20 minutes, by the end of which all of the sirolimushad gone into solution to form a sirolimus stock solution. A diluentsolvent was prepared by sonicating about 1880 g of PEG 400 for about 60minutes, and then sparging the solvent with nitrogen for about 10minutes.

The sirolimus stock solution and the PEG 400 were then rotated at roomtemperature in a rotary evaporator for about 10 minutes to mix the stocksolution with the diluent solvent. After mixing, the solution wassparged with nitrogen for about 10 minutes and blanketed with nitrogenfor about 5 minutes. After the solution was sparged and filled withnitrogen, about 240 g of excess ethanol was evaporated from the solutionby increasing the solution temperature, maintaining a temperature thatdid not exceed 40° C. for an extended period of time and continuing torotate the solution for about 2.5 hours.

The resulting solution comprised about 40 g of sirolimus (about 2% byweight), about 80 g of ethanol (about 4% by weight), and about 1880 g ofPEG 400 (about 94% by weight). This solution was sparged with nitrogenfor about 10 minutes and blanketed with nitrogen for about 5 minutes.The solution was then filtered through a 0.2 micron filter. USP type Iglass vials were filled with 0.5 ml each of the filtered solution toleave a head space in each container of about 2 ml. This head space wasfilled with nitrogen gas and capped. This solution is listed asformulation #340 in Table 1.

Example 47 Preparation and Characterization of a Rapamycin-ContainingSolution

About 32 g of ethanol was sparged with N₂ for about 10 minutes, and thenabout 4 g of sirolimus was added to the ethanol. The mixture wassonicated for about 20 minutes, by the end of which all of the sirolimushad gone into solution to form a sirolimus stock solution. A diluentsolvent was prepared by sonicating about 92 g of PEG 400 for about 60minutes, and then sparging the solvent with Nitrogen for about 10minutes.

The sirolimus stock solution and the PEG 400 were then rotated at aboutroom temperature in a rotary evaporator for about 10 minutes to mix thestock solution with the diluent solvent. After mixing, the solution wassparged with nitrogen for about 10 minutes and blanketed with nitrogenfor about 5 minutes. After the solution was sparged and filled withnitrogen, about 28 g of excess ethanol was evaporated from the solutionby increasing the solution temperature, maintaining a temperature thatdid not exceed 40° C. for an extended period of time and continuing torotate the solution for about 2.5 hours.

The resulting solution comprised about 4 g of sirolimus (about 4% byweight), about 4 g of ethanol (about 4% by weight), and about 92 g ofPEG 400 (about 92% by weight). This solution was sparged with nitrogenfor about 10 minutes and blanketed with nitrogen for about 5 minutes.The solution was then filtered through a 0.2 micron filter. USP type Iglass vials were filled with 0.5 ml each of the filtered solution toleave a head space in each container of about 2 ml. This head space wasfilled with nitrogen gas and capped. This solution is listed asformulation #326 in Table 1.

Example 48 Preparation and Characterization of a Rapamycin-ContainingSolution

About 200 μL of normal saline (sodium chloride, 0.9%) was added to 0.5mL of the solution of Example 46 (previously filled in 2 mL USP type Iglass vials) using a 0.5 cc insulin syringe, and mixed by gentlyswirling. The resulting mixture is a clear solution and has anapproximate composition of 1.47% (w/w) of sirolimus, 2.93% (w/w) ofethanol, 26.6% (w/w) of normal saline, and 69% (w/w) of PEG 400. Thissolution is listed as formulation #327 in Table 1.

Example 49 Preparation and Characterization of a Rapamycin-ContainingSolution

About 250 μL of normal saline (sodium chloride, 0.9%) was added to 0.5mL of the solution of Example 46 (previously filled in 2 mL USP type Iglass vials) using a 0.5 cc insulin syringe, and mixed by gentlyswirling. The resulting mixture is a dispersed form and has anapproximate composition of 1.38% (w/w) of sirolimus, 2.75% (w/w) ofethanol, 31.25% (w/w) of normal saline, and 64.62% (w/w) of PEG 400.This solution is listed as formulation #328 in Table 1.

Example 50 Subconjunctival Injection of a Rapamycin-Containing Solution

Either 40 μl of the solution described in Example 46, Example 48, orExample 49, or 20 μL of the solution described in Example 47 wasinjected between the sclera and the conjunctiva of the eye of NewZealand White rabbits. FIG. 1 depicts the average concentration ofrapamycin present in the retina choroid (ng/g) on a logarithmic scale at14, 30, 60, and 90 days after injection.

The analysis was by liquid chromatography mass spectroscopy (LCMS) usingan internal standard. At each time point, the average concentration ofrapamycin was calculated by adding the concentrations of rapamycinobtained for each eye from each rabbit, and dividing the total by thenumber of eyes analyzed. In this experiment, each time point representsthe average of either two eyes of each of two rabbits (four eyes at thattime point).

The full retina choroid was homogenized and analyzed. The averageconcentration of the retina choroid was calculated by dividing the massof rapamycin measured by the mass of retina choroid analyzed. The sampledid not include the site of administration; thus, this measurementindicated the level of rapamycin delivered to the retina choroid viatest article (in this case, the solutions of Examples 46, 47, 48, and49).

The average level of rapamycin in the retina choroid at 14, 30, 60, and90 days after subconjunctival injection of the solution of Example 46was about 56.94, 9.79, 2.26, and 0.00 ng/g, respectively. The averagelevel of rapamycin in the retina choroid at 14, 30, 60, and 90 daysafter subconjunctival injection of the solution of Example 47 was about90.19, 6.0, 5.72, and 0.98 ng/g, respectively. The average level ofrapamycin in the retina choroid at 14, 30, 60, and 90 days aftersubconjunctival injection of the solution of Example 48 was about103.86, 20.54, 6.07, and 12.76 ng/g, respectively. The average level ofrapamycin in the retina choroid at 14, 30, 60, and 90 days aftersubconjunctival injection of the solution of Example 49 was about 70.12,27.83, 3.99, and 5.94 ng/g, respectively.

Example 51 Intravitreal Injection of a Rapamycin-Containing Solution

6 μl of the solution of Example 46 or 8.5 μL of the solution of Example48 was injected into the vitreous of the eye of New Zealand Whiterabbits using a 10 μL Hamilton syringe. FIG. 2 depicts the level ofrapamycin present in the retina choroid (ng/g) on a logarithmic scale at3, 14, 30, 45 and 60 days after injection.

Full vitreous of each eye was homogenized and analyzed by liquidchromatography tandem mass spectrometry (LC/MS/MS). The average level ofrapamycin in the vitreous at 3, 14, 30 and 60 days after intravitrealinjection of the solution of Example 46 was about 63,064.3, 6,095.56,58.21, and 0.15 ng/ml, respectively. The average level of rapamycin inthe vitreous at 3, 14, 30, 45 and 60 days after intravitreal injectionof the solution of Example 48 was about 58,930.4, 18,573.1, 6,793.21,799.18, and 4116.17 ng/ml, respectively.

The full retina choroid was homogenized and analyzed. The averageconcentration of the retina choroid was calculated by dividing the massof rapamycin measured by the mass of retina choroid analyzed. The sampledid not include the site of administration; thus, this measurementindicated the level of rapamycin delivered to the retina choroid viatest article (in this case, the solution of Example 46 or 48). Theaverage level of rapamycin in the retina choroid at 3, 14, 30, and 60days after intravitreal injection of the solution of Example 46 wasabout 1971.84, 478.02, 57.70, and 1.90 ng/g, respectively. The averagelevel of rapamycin in the retina choroid at 3, 14, 30, 45 and 60 daysafter intravitreal injection of the solution of Example 48 was about2869.05, 3633.64, 43.38, 37.09 and 27.36 ng/g, respectively.

Example 52 Intravitreal and Subtenon Injection of a Rapamycin-ContainingSolution

30 μl of the solution of Example 46 was injected between the sclera andthe conjunctiva of the eye of New Zealand White rabbits. FIG. 3 depictsthe average concentration of rapamycin present in the retina choroid(ng/g) and whole blood (ng/mL) on a logarithmic scale at 3, 7, 14, 30,60 and 90 days after injection.

The full retina choroid was processed and analyzed as described earlier.The average level of rapamycin in the retina choroid at 3, 7, 14, 30, 60and 90 days after subconjunctival injection of the solution of Example46 was about 174.1, 129.5, 34.7, 7.8, 1.7, and 1.7 ng/g, respectively.The whole blood samples are precipitated by addition of known amountorganic solvents, homogenized, centrifuged, and supernatant wascollected to analyze rapamycin levels using LC/MS/MS. The average levelof rapamycin in the blood at 3, 7, 14, 30 and 60 days aftersubconjunctival injection of the solution of Example 46 was about 13.0,4.6, 2.0, 0.3, and 0.3 ng/mL, respectively.

For the 30 μL subtenon injections, a guide hole was made by insertingand removing a 20 gauge×½ inch sharp BD needle though the conjunctiva inthe dorsotemporal quadrant 3 mm posterior to the limbus and temporal tothe dorsal rectus muscle attachment. A 24 G fukasaku anesthesia cannulaattached to a sterile 0.5 cc BD syringe was then guided through theguide hole posteriorly into the subtenon space and the solution ofExample 46 was injected slowly on the posterior surface of the scleranear the optic nerve. FIG. 3 depicts the average concentration ofrapamycin present in the retina choroid (ng/g) and whole blood (ng/mL)on a logarithmic scale at 3, 7, 14, 30, 60 and 90 days after injection.

The full retina choroid was processed and analyzed as described earlier.The average level of rapamycin in the retina choroid at 3, 7, 14, 30, 60and 90 days after subtenon injection of the solution of Example 46 wasabout 202.1, 49.6, 10.5, 6.0, 1.7, and 3.9 ng/g, respectively. The wholeblood samples are precipitated by addition of known amount organicsolvents, homogenized, centrifuged, and supernatant was collected toanalyze rapamycin levels using LC/MS/MS. The average level of rapamycinin the blood at 3, 7, 14, 30 and 60 days after subtenon injection of thesolution of Example 46 was about 23.8, 5.9, 2.1, 0.4, and 0.8 ng/mL,respectively.

Example 53 Preparation and Characterization of a Rapamycin-ContainingSolution

The following solvent preparation allows up to 5% (w/w) of rapamycin.N-methyl pyrrolidone (NMP) and propylene glycol (PG) are mixed in10.5:73.7 ratio, desired amount of sirolimus (up to 5% w/w of finalcomposition) is dissolved in the mix, and subsequently water (up to15.8% w/w of final composition) is added slowly to obtain the finalsolution. This solution is listed as formulation #333 in Table 1.

Example 54 Preparation and Characterization of a Rapamycin-ContainingSolution

The following solvent preparation allows up to 5% (w/w) of rapamycin.N-methyl pyrrolidone (NMP) and propylene glycol (PG) are mixed in38.9:38.9 (1:1) ratio, desired amount of sirolimus (up to 5% w/w offinal composition) is dissolved in the mix, and subsequently water (upto 22.2% w/w of final composition) is added slowly to obtain the finalsolution. This solution is listed as formulation #334 in Table 1.

Example 55 Preparation and Characterization of a Rapamycin-ContainingSolution

The following solvent preparation allows up to 5% (w/w) of rapamycin.N-methyl pyrrolidone (NMP) and polyethylene glycol 600 (PEG 600) aremixed in 10.5:73.7 ratio, desired amount of sirolimus (up to 5% w/w offinal composition) is dissolved in the mix, and subsequently water (upto 15.8% w/w of final composition) is added slowly to obtain the finalsolution. This solution is listed as formulation #335 in Table 1.

Example 56 Preparation and Characterization of a Rapamycin-ContainingSolution

The following solvent preparation allows up to 5% (w/w) of rapamycin.Dimethyl acetamide (DMA) and propylene glycol (PG) are mixed in10.5:73.7 ratio, desired amount of sirolimus (up to 5% w/w of finalcomposition) is dissolved in the mix, and subsequently water (up to15.8% w/w of final composition) is added slowly to obtain the finalsolution. This solution is listed as formulation #336 in Table 1.

Example 57 Preparation and Characterization of a Rapamycin-ContainingSolution

The following solvent preparation allows up to 5% (w/w) of rapamycin.Dimethyl sulfoxide (DMSO) and propylene glycol (PG) are mixed in10.5:73.7 ratio, desired amount of sirolimus (up to 5% w/w of finalcomposition) is dissolved in the mix, and subsequently water (up to15.8% w/w of final composition) is added slowly to obtain the finalsolution. This solution is listed as formulation #337 in Table 1.

Example 58 Preparation and Characterization of a Rapamycin-ContainingSolution

N-methylpyrrolidone (NMP), polyethylene glycol 400 (PEG 400) and ethanolare mixed in 27.9:28.4:4.7 ratio (respectively), desired amount ofsirolimus (up to 5% w/w of final composition) is dissolved in the mix,and subsequently water (up to 28.4% w/w of final composition) is addedslowly to obtain the final solution. This solution is listed asformulation #338 in Table 1.

Example 59 Preparation and Characterization of a Rapamycin-ContainingSolution

8.35% of Cremophor (percentage of total weight) is mixed with 8.37% ofCapmul MCM (percentage of total weight). 0.07% rapamycin (percentage ofthe total weight) was dissolved in the Cremophor-Capmul MCM solutionusing an ultrasound bath. 83.21% (percentage of total weight) of sterilewater is added to the former solution and the resulting solution isvortexed creating a milky microemulsion. The solution was placed at 2-8°C. until use. This solution is listed as formulation #329 in Table 1.

Example 60 Preparation and Characterization of a Rapamycin-ContainingSolution

8.15% of Capmul MCM (percentage of total weight) is mixed with 8.12% ofTyloxapol (percentage of total weight). 0.09% rapamycin (percentage ofthe total weight) was dissolved in the Tyloxapol-Capmul MCM solution inwhich 0.35% of Ethanol was added. The final dissolution is completed byusing an ultrasound bath. 83.29% (percentage of total weight) of sterilewater is added to the former solution and the resulting solution isvortexed creating a milky microemulsion. The solution was placed at 2-8°C. until use. This solution is listed as formulation #330 in Table 1.

Example 61 Preparation and Characterization of a Rapamycin-ContainingSolution

6.81% of Tyloxapol (percentage of total weight) is mixed with 6.81% ofCapmul MCM (percentage of total weight) and 3.12% of PHOSAL® 50PG(percentage of total weight). 0.08% rapamycin (percentage of the totalweight) was dissolved in the Tyloxapol-Capmul MCM-PHOSAL® 50PG solutionusing an ultrasound bath. 83.21% (percentage of total weight) of sterilewater is added to the former solution and the resulting solution isvortexed creating a milky microemulsion. The solution was placed at 2-8°C. until use. This solution is listed as formulation #331 in Table 1.

Example 62 Preparation and Characterization of a Rapamycin-ContainingSolution

7.18% of Cremophor (percentage of total weight) is mixed with 9.47% ofCapmul MCM (percentage of total weight). 0.17% rapamycin (percentage ofthe total weight) was dissolved in the Cremophor-Capmul MCM solution inwhich 0.35% of Ethanol was added. 83.18% (percentage of total weight) ofsterile water was added to the former solution and the resultingsolution is vortexed creating a milky microemulsion. The solution wasplaced at 2-8° C. until use. This solution is listed as formulation #332in Table 1.

Example 63 Topical Administration of Rapamycin-Containing Formulations

Both eyes (or one eye depending on the study) of each animal receivedone or two drops (40 μl per drop) of the studied formulations using acalibrated pipette on day 0 (day of dosing). Results are shown in Tables3-6 and FIGS. 4-9.

For Tables 3-6, the solution of Example 46 (formulation #340) wasadministered two drops twice a day on both eyes, one time for Group I,two drops once a day on one eye, one time for Group II, and two dropsonce a day on both eyes for 6 days in Group III. In Group II, the righteye (OD) but not the left eye (OS) received eye drops. In a furtherstudy, one drop of the formulation #340 (40 μL containing 880 μgrapamycin) was administered on day zero. As shown in FIG. 4, afteradministration of a single eye drop, the rapamycin concentration in thecornea remained above 0.01 ng/g nine days (e.g., 216 hrs) afteradministration, while the rapamycin concentration in retina choroidremained above 0.001 ng/g nine days (e.g., 216 hrs) afteradministration. In fact after administration of a single eye drop, therapamycin concentration in the cornea remained above 10.0 ng/g (above0.01 ng/mg) nine days (e.g., 216 hrs) after administration, while therapamycin concentration in retina choroid remained above 1.0 ng/g (above0.001 ng/mg) nine days (e.g., 216 hrs) after administration.

Moreover as shown in FIG. 9 in a further study, following administrationof a single non-aqueous eye drop formulation containing 4%, 1% or 0.5%rapamycin, sustained levels of rapamycin were detected in the cornea andretina choroid. In particular, after administration of a single eyedrop, the rapamycin concentration in the cornea remained above 0.01 ng/g(above 0.01 ng/mg) for at least nine days after administration, whilethe rapamycin concentration in retina choroid remained above 0.001 ng/g(above 0.001 ng/mg) for at least 9 days after administration. Incontrast, the rapamycin concentration in the blood was substantiallylower than that observed in the cornea and retina choroid indicatingthat the non-aqueous eye drop formulations did not result in significantsystemic exposure to rapamycin.

For FIGS. 5 and 6, the aqueous rapamycin-containing formulations #329and #330 of Table 1, were administered two drops once or twice a day forup to 28 days. For FIGS. 7 and 8, the aqueous rapamycin-containingformulations #331 and #332 of Table 1 were administered two drops twicea day for 6 days. In short, the aqueous rapamycin-containingformulations tested achieved an average concentration of rapamycin inthe cornea of at least 0.01 ng/g (greater than 0.01 ng/mg to 0.1 ng/mgin FIGS. 5-8), in the retina choroid of at least 0.01 ng/g (greater than0.001 ng/mg to 0.01 ng/mg in some measurements of FIGS. 5-8), in theaqueous humor of at least 0.1 ng/mL (greater than 1.0 ng/mL), and in thevitreous humor of at least 0.1 ng/mL (greater than 1.0 ng/mL).

For FIGS. 10A and 10B one drop of an aqueous or a non-aqueousformulation containing about 0.2% rapamycin were administered to botheyes of rabbit subjects once a day for six days (day 0 to day 6). Asused herein, the terms “aqueous” and “hydrophilic” refer to formulationsin which the dominant component is an aqueous compound such as water(e.g., formulations #342 and #342 of Table 1), while the terms“non-aqueous” and “lipophilic” refer to formulation in which thedominant component is a non-aqueous compound such as PEG 400 (e.g.,formulations #344 and #345 of Table 1). Rapamycin concentrations in thecornea, retina choroid and blood were measured at day 7 and day 14. Bothformulations tested achieved an average concentration of rapamycin inthe cornea of at least 0.01 ng/g (at least 10 ng/g) on days 7 and 14,and in the retina choroid of at least 0.01 ng/g (at least 1 ng/g) ondays 7 and 14.

For FIG. 11 one drop of a non-aqueous formulation containing about 0.2%rapamycin (e.g., formulations #344 or #345) was administered to botheyes of rabbit subjects once a day on day 0, 7, 14 and 21. Rapamycinconcentrations in the cornea, retina choroid and aqueous humor weremeasured on days 3, 10, 17, 24 and 28. Infrequent administration of thenon-aqueous eye drops achieved sustained therapeutic levels ofrapamycin. In particular a sustained rapamycin concentration wasachieved in the cornea of at least 0.01 ng/g (above 10 ng/g) and in theretina choroid and aqueous humor of at least 0.01 ng/mL (above 1 ng/mL).

Example 64 Preparation and Intravitreal Administration of aRapamycin-Containing Formulation

An exemplary rapamycin containing formulation was prepared by adding0.2003 g rapamycin to 5.9989 g NMP. After mixing, 7.8030 g PG is added,followed by 6.010 g sterile water. This recipe yielded a formulationhaving final concentrations as a percentage of the total weight ofapproximately: rapamycin 1.0%, NMP 30.0%, PG 39.0% and sterile water30.0%. This formulation is listed as #346 in Table 1.

The formulation of this example, however, was not stable. As such at thetime of the in vivo study (ten days after this preparation was made) therapamycin potency was determined to be 0.6%. Twenty μl of the 0.6%rapamycin-containing formulation was injected into the vitreous of theeye of New Zealand White rabbits. Rapamycin concentrations in vitreous,retina choroid and sclera were determined as described in Example 51.FIG. 12 depicts the level of rapamycin present in the vitreous, retinachoroid and sclera 30 days or more (e.g., 30, 60, 90 and 120 days ormore) after injection.

Example 65 Preparation and Subconjunctival Administration of RapamycinFormulations

An exemplary 2% rapamycin suspension was prepared by dispersing 106.9 mgof rapamycin (2.07% by weight) in 5056.3 mg of 1% carboxymethylcellulose(CMC) low viscosity (97.93% by weight) aqueous solution. Briefly, 106.9mg of rapamycin and 5056.3 mg of 1% carboxymethylcellulose low viscositywere placed in an amber vial. High wear resistant Ziconia grinding media(beads) of 3 mm diameter were added, up to ¾ of the total volume. Thevial was sealed and placed in a Cole-Parmer milling apparatus for atleast 24 hrs. The median and mean particle sizes for rapamycin were3.2957 μm and 3.5487 μm, respectively. The formulation was kept at 4° C.until use. Final concentrations as a percentage of the total weight wereapproximately: rapamycin: 2.07%, CMC 1%, and sterile water 97.93%. Thisformulation is listed as #347 in Table 1.

Another exemplary 2% rapamycin in situ gelling formulation was preparedby dissolving 459.9 mg of PLGA (75/25) in 1678.0 mg of NMP overnight.Then 44.8 mg of rapamycin is added until complete dissolution. Theformulation was kept at 4° C. until use. Final concentrations as apercentage of the total weight were approximately: rapamycin 2.05%, PLGA75/25 21.07%, and NMP 76.88%. This formulation is listed as #348 inTable 1.

A further exemplary 2% rapamycin formulation was prepared by dissolvingrapamycin in NMP. Then PEG 400 is added under continuous agitation,followed by sterile water also under continuous agitation. Theformulation was kept at 4° C. until use. Final concentrations as apercentage of the total weight were approximately: rapamycin 2.02%, NMP44.47%, PEG 400 29.04%, and sterile water 24.47%. This formulation islisted as #349 in Table 1.

Thirty μl of the 2% rapamycin-containing formulations are injectedbetween the sclera and the conjunctiva of the eye of New Zealand Whiterabbits. Rapamycin concentrations in vitreous, retina/choroid and scleraare determined as described in Example 50, at days 30, 60 and 90 dayspost-administration.

Example 66 Preparation and Topical Administration of aRapamycin-Containing SDF

An exemplary rapamycin-containing solid dosage form (SDF) was preparedusing a solvent cast technique. Briefly, the following components weremixed together: 10.0% rapamycin, 57.9% KOLLIDON 90F (PVP 90), 20.0%phosal 50 SD, 10.1% PEG 400, 1.0% vitamin E (gamma tocopherol), and 1.0%ascorbyl palmitate (e.g., equivalent to the formulation #351 in Table1). A volume of ethanol was added sufficient to obtain 30% total solids(e.g., 70% ethanol) as a final percentage. The mixture was then pouredonto a release liner (polyester film with a silicone release coating,and spread to a thickness of 50 mm using a gardner knife. The ethanolwas evaporated off in a dark, nitrogen rich environment (i.e., glovebox) for about 24 hrs. After drying, the concentration of rapamycin inthe SDF was determined by liquid chromatography to be 9.1±0.1% (e.g.,0.9% rapamycin lost). The dried formulation was punched to the desireddiameter. The SDF of this example had a diameter of 4 mm, with athickness of 0.15-0.20 mm, yielding a rapamycin dosage of about 246±21μg.

The SDF was administered topically to the eyes of New Zealand Whiterabbits by placement in the inferior cul-de-sac. Briefly, the lower lidof the eye was pulled out, permitting the placement at the bottom of thecul-de-sac on the inferior side (not sclera side) with a pair oftweezers. Rapamycin concentrations in cornea, retina choroid, vitreoushumor, aqueous humor and blood were determined. FIG. 13 depicts thelevels of rapamycin in various tissues on days 1, 7, 14, 21 and 28post-administration.

Example 67 Preparation of a Dexamethasone-Containing SDF

An exemplary dexamethasone-containing solid dosage form (SDF) isprepared using a solvent cast technique. The SDF is prepared withconcentrations as a percentage of the total weight of approximately:25.3% dexamethasone, 33.2% hydroxypropyl cellulose (HPC), 32.0% eudragitRS 30D, and 9.5% xanthan gum. The SDF of less than or equal to 3 mm indiameter is inserted subconjunctivally, for controlled release in theretina/choroid for up to six months (1 to 100 ng/g dexamethasone). Thisformulation is listed as #352 in Table 1.

Example 68 Preparation and Invitreal Administration of a DasatanibFormulation

An exemplary dasatanib-containing solution was prepared by dissolvingdasatanib in DMA. PEG 400 was then added under continuous agitation.Final concentrations as a percentage of the total weight wereapproximately: dasatanib 4.8%, dimethylacetamide 16.0%, and PEG 40079.2% (e.g., equivalent to the formulation #339 in Table 1).

Twenty μl of the 4.84% dasatanib-containing formulation was injectedinto the vitreous of the eye of New Zealand White rabbits. Dasatanibconcentrations in the vitreous and retina choroid were determined. FIG.14 depicts the level of dasatanib present in the vitreous and retinachoroid about 2 weeks, 5 weeks and 10 weeks after injection.

TABLE 1 Formulations Median NDM, Form. # Composition (mg), % (w/w) Typeparticle size Injection volume 1 DMSO = 2000 mg (20%) S Water = 8000 mg(80%) 2 F68 = 1000 mg (10%) S Water = 9000 mg (90%) 3 F68 = 3000 mg(30%) S Water = 7000 mg (70%) 4 F127 = 1000 mg (10%) S Water = 9000 mg(90%) 5 F127 = 1500 mg (15%) S Water = 8500 mg (85%) 6 Beta-cyclodextrin= 250 mg (2.5%) S Water = 9750 mg (97.5%) 7 Rapa = 10.2 mg (0.101%) SNo, 50 μL Pluronic, F68 = 1010 mg (9.99%) Water = 9090 mg (89.909%) 8Rapa = 10.2 mg (0.102%) S No, 50 μL Pluronic, F68 = 3000 mg (29.969%)Water = 7000 mg (69.929%) 9 Rapa = 10.5 mg (0.104%) S No, 50 μLPluronic, F127 = 1010 mg (9.99%) Water = 9090 mg (89.907%) 10 Rapa =10.5 mg (0.105%) S No, 50 μL Pluronic, F127 = 1500 mg (14.984%) Water =8925 mg (84.9%) 11 Rapa = 10.7 mg (0.105%) S No, 50 μL Beta-cyclodextrin= 255 mg (2.497%) Water = 9945 mg (97.398%) 12 Rapa = 6.4 mg (0.0999%)SP CMC = 48 mg (0.7493%) Polysorbitan 20 = 2.56 mg (0.04%) Water =6349.44 mg (99.111%) 13 Rapa = 6.5 mg (0.0999%) S DMSO = 325 mg (4.995%)Water = 6175 mg (94.905%) 14 Rapa = 13.5 mg (0.0999%) SP CMC = 101.25 mg(0.7493%) Polysorbitan 20 = 5.4 mg (0.04%) Water = 13393.35 mg (99.112%)15 Rapa = 11.0 mg (0.2%) S EtOH = 5500 mg (99.8%) 16 Rapa = 6.6 mg(0.1%) S EtOH = 1054.6 mg (15.933%) F127 = 833.64 mg (12.595%) Water =4723.96 mg (71.372%) 17 Rapa = 5 mg (0.1%) S Cavitron = 0.25 g (5%)Ethanol, 95% = 57 mg (1.1%) Sterile water = 4.753 g (93.8%) 18 Rapa = 5mg (0.1%) S Ethanol, 95% = 150 mg (2.9%) PEG400 = 1.0 g (19.4%) Sterilewater = 4.01 g (77.6%) 19 Rapa = 5 mg (0.1%) S Yes, 50 μL Ethanol, 95% =152 mg (3.2%) PEG400 = 1.5227 g (30.2%) Sterile water = 3.3592 g(66.67%) 20 Rapa = 6.6 mg (0.1%) S EtOH = 505.1 mg (7.618%) F127 = 917.8mg (13.843%) Water = 5200.6 mg (78.44%) 21 Rapa = 6.6 mg (0.1%) S No, 50μL EtOH = 536 mg (7.5%) Pluronic, F127 = 983.75 mg (14.0%) Water =5574.56 mg (78.4%) 22 Rapa = 5.2 mg (0.1023%) S EtOH = 56.6 mg (1.127%)Captisol = 2008.9 mg (39.5%) Water = 3013.3 mg (59.3%) 23 Rapa = 6.9 mg(0.201%) S EtOH = 3418.0 mg (99.799%) 24 Rapa = 9.1 mg (0.491%) S EtOH =90.9 mg (4.908%) F127 = 262.8 mg (14.191%) Water = 1489.1 mg (80.409%)25 Rapa = 0 mg (0%) S EtOH = 310.2 mg (5.144%) F127 = 858.1 mg (14.228%)Water = 4862.6 mg (80.628%) 26 Rapa = 0 mg (0%) S EtOH = 613.1 mg(10.19%) F127 = 810.6 mg (13.471%) Water = 4593.6 mg (76.339%) 27 Rapa =53.5 mg (1.095%) S Yes, 50 μL EtOH = 414.8 mg (8.488%) F127 = 662.8 mg(13.563%) Water = 3755.7 mg (76.854%) 28 Rapa = 0.3 g (10%) ISG, PVP K90= 0.35 g (12%) SP Eudragit RS30D = 2.35 g (78%) 29 Rapa = 0.2154 g(7.31%) ISG, PVP K90 = 0.25 g (8.5%) SP Eudragit RS30D = 2.48 g (84.19%)30 Rapa = 53.9 mg (1.103%) S No, 50 μL EtOH = 413.6 mg (8.463%) Sterilewater = 3843.5 mg (78.647%) F127 (Lutrol) = 576.0 mg (11.786%) 31 Rapa =0 mg (0%) S EtOH = 411.9 mg (8.513%) Sterile Water = 3849.3 mg (79.554%)F127(Lutrol) = 577.4 mg (11.933%) 32 Rapa = 54.1 mg (1.256%) S EtOH =416.8 mg (9.676%) Sterile Water = 3836.3 mg (78.569%) F127(Lutrol) =577.5 mg (10.499%) 33 Rapa = 80.7 g (1.964%) S EtOH = 65.0 mg (0.158%)PEG400 = 4021.8 mg (97.878%) 34 Rapa = 106.9 g (5.233%) S Yes, 25 μLEtOH = 129.6 mg (6.344%) PEG400 = 1806.5 mg (88.424%) 35 Rapa = 0 mg(0%) ISG, PVP K90 = 0.204 g (2.3%) SP Ethanol, 100% = 0.4 g (4.5%)Eudragit RL100 = 0.201 g (2.3%) PEG 400 = 8.00 g (90.9%) 36 Rapa = 0 mg(0%) ISG, PVP K90 = 0.2 g (2.2%) SP Ethanol, 100% = 0.4 g (4.4%) PVAP =0.4 g (4.4%) PEG 400 = 8.00 g (88.9%) 37 Rapa = 106.1 mg (4.2%) ISG, PVPK90 = 55.2 mg (2.2%) SP Ethanol, 100% = 108 mg (4.3%) Eudragit RL100 =55 mg (2.2%) PEG 400 = 2.2 g (87.1%) 38 Rapa = 399.6 mg (9.965%) S Yes,20 μL F68(Lutrol) = 40.6 mg (1.012%) Sterile Water = 3569.7 mg (89.022%)39 Rapa = 53.8 mg (1.1%) S EtOH = 415.2 mg (8.489%) Sterile Water =3844.2 mg (78.594%) F127 = 578.0 mg (11.817%) 40 Rapa = 208.1 mg(3.148%) S Yes, 20 μL PEG400 = 6403.4 mg (96.852%) 41 Rapa = 200.4 mg(5.148%) SP F68(Lutrol) = 20.8 mg (0.534%) PEG400 = 3569.3 mg (91.697%)EtOH (95%) = 102 mg (2.62%) 42 Rapa = 200.4 g (5.259%) SP PEG400 =3561.4 mg (93.46%) Tween 80 = 48.8 mg (1.281%) 43 Rapa = 30.9 mg (1.03%)S No, 50 μL PEG 400 = 2.9624 g (98.97%) 44 Rapa = 61 mg (1.96%) S Yes,50 μL Ethanol, 100% = 0.1860 g (6%) PEG 400 = 2.8588 g (92.04%) 45 Rapa= 90.7 mg (3.02%) S Yes, 50 μL Ethanol, 100% = 0.2722 g (9.06%) PEG 400= 2.6423 g (87.94%) 46 Rapa = 101.6 mg (4.997%) S EtOH = 331.6 mg(16.308%) PEG400 = 1600.1 mg (78.695%) 47 Rapa = 120.9 g (3.189%) SPF68(Lutrol) = 42.4 mg (1.118%) Sterile Water = 3627.7 mg (95.692%) 48Rapa = 100.1 g (1.999%) S EtOH = 305.1 mg (6.092%) PEG400 = 4602.9 mg(91.909%) 49 Rapa = 150.5 mg (3.004%) SP Yes, 20 μL, 40 μL PEG400 =4860.3 mg (96.996%) 50 Rapa = 153.4 mg (3.055%) SP No, 20 μLF68(Pluronic) = 50.6 mg (1.008%) Sterile Water = 4816.6 mg (95.937%) 51Rapa = 116.6 mg (2.29%) S Yes, 30 μL EtOH = 306.6 mg (6.05%) PEG400 =4647.5 mg (91.66%) 52 Rapa = 150.4 mg (2.994%) SP F68 Lutrol = 15.4 mg(0.306%) Sterile water = 4859.1 mg (96.7%) 53 Rapa = 306.5 mg (6.088%)SP PEG 400 = 4727.7 mg (93.912%) 54 Rapa = 309.3 mg (6.146%) SP PEG 400= 4723.3 mg (93.854%) 55 Rapa = 303.3 mg (6.061%) SP PEG 400 = 4700.6 mg(93.939%) 56 Rapa = 305.4 mg (6.088%) SP PEG 400 = 4711.0 mg (93.912%)57 Rapa = 306.9 mg (6.098%) SP PEG 400 = 4725.5 mg (93.902%) 58 Rapa =302.5 mg (6.021%) SP PEG 400 = 4721.6 mg (93.979%) 59 Rapa = 304.5 mg(6.053%) SP PEG 400 = 4726.4 mg (93.947%) 60 Dexamethasone = 251.4 mg(5.011%) SP PEG 400 = 4765.2 mg (94.989%) 61 Dexamethasone = 252.4 mg(5%) SP PEG 400 = 4600 mg (92%) EtOH = 150 mg (3%) 62 Rapa = 32.2 mg(0.641%) S PEG 400 = 4677.9 mg (93.096%) EtOH = 314.7 mg (6.263%) 63Rapa = 32.3 mg (0.6%) S PEG 400 = 5516.3 mg (93.1%) EtOH = 314.7 mg(6.263%) 64 Rapa = 54.4 mg (1.007%) S PEG 400 = 4638.9 mg (92.702%) EtOH= 314.8 mg (6.291%) 65 Rapa = 50.8 mg (1.013%) S PEG 400 = 4963.2 mg(98.987%) 66 Rapa = 52.1 mg (1.035%) S PEG 400 = 4868.6 mg (96.718%)EtOH = 113.1 mg (2.247%) 67 Rapa = 50.5 mg (1.009%) S Yes, 20 μL PEG 400= 4752.8 mg (94.953%) No, 40 μL, 100 μL EtOH = 202.1 mg (4.038%) 68 Rapa= 101.8 mg (2.030%) S PEG 400 = 4712.4 mg (93.970%) EtOH = 200.6 mg(4.000%) 69 Rapa = 102.1 mg (2.036%) S PEG 400 = 4605.5 mg (91.847%)EtOH = 306.7 mg (6.117%) 70 Rapa = 101.6 mg (2.025%) S PEG 400 = 4510.6mg (89.892%) EtOH = 405.6 mg (8.083%) 71 Rapa = 75.9 mg (3.019%) SP PEG400 = 2438.4 mg (96.981%) 72 Rapa = 50.9 mg (2.034%) S PEG 400 = 2350.1mg (93.914%) EtOH = 101.4 mg (4.052%) 73 Rapa = 12.5 mg (0.620%) SP PEG400 = 2004.8 mg (99.380%) 74 Rapa = 1.20949 g (2.0152%) S EtOH = 2.401 g(4.000%) PEG 400 = 56.407 g (93.9848%) 75 Rapa = 16.0 mg g (0.795%) SNo, 50 μL EtOH = 80.0 mg (3.976%) PEG 400 = 1916.0 mg (95.2298%) 76 Rapa= 8.1 mg (0.400%) SP PEG 400 = 2014.5 mg (99.600%) 77 Rapa = 8.6 mg(0.428%) S PEG 400 = 2002.5 mg (99.572%) 78 Rapa = 8.2 mg (0.410%) S PEG400 = 1992.0 mg (99.590%) 79 Rapa = 8.7 mg (0.433%) S PEG 400 = 1998.8mg (99.567%) 80 Rapa = 8.6 mg (0.427%) S PEG 400 = 2003.2 mg (99.573%)81 Rapa = 8.6 mg (0.428%) S PEG 400 = 1999.3 mg (99.572%) 82 Rapa = 9.0mg (0.448%) S PEG 400 = 2000.8 mg (99.552%) 83 Rapa = 8.0 mg (0.397%) SPEG 400 = 2008.8 mg (99.603%) 84 Rapa = 8.5 mg (0.422%) S PEG 400 =2006.8 mg (99.578%) 85 Rapa = 8.0 mg (0.399%) S PEG 400 = 1998.2 mg(99.601%) 86 Rapa = 8.5 mg (0.422%) S PEG 400 = 2004.3 mg (99.578%) 87Rapa = 8.6 mg (0.428%) S PEG 400 = 2002.5 mg (99.572%) 88 Rapa = 0.7 g(1.983%) S EtOH = 1.4 g (3.966%) PEG 400 = 33.2 g (94.051%) 89 Rapa = 0g (0%) S EtOH = 0.574 g (1.995%) PEG 400 = 28.2 g (98.005%) 90 Rapa =1.95 g (1.950%) S EtOH = 4.05 g (4.050%) PEG 400 = 94.00 g (94.000%) 91Rapa = 0.0107 g (0.534%) S No, 80 μL EtOH = 0.0805 g (4.019%) PEG 400 =1.912 g (95.447%) 92 Rapa = 0.0081 g (0.403%) S No, 100 μL EtOH = 0.0804g (4.003%) PEG 400 = 1.920 g (95.594%) 93 Rapa = 1.992 g (2%) S EtOH =3.9419 (4%) PEG 400 = 93.95 g (94%) 94 Rapa = 0.405 g (0.4%) S EtOH =4.24 g (4%) PEG 400 = 95.6 (95.6%) 95 PEG 400 = 96 g (96%) S EtOH =3.9027 (4%) 96 Rapa = 0.4020 g (0.402%) S EtOH = 3.970 g (3.971%) PEG400 = 95.600 g (95.627%) 97 Rapa = 2.000 g (1.990%) S EtOH = 4.000 g(3.980%) PEG 400 = 94.500 g (94.030%) 98 PEG 400 = 96 g (96%) S EtOH =3.92 g (4%) 99 Rapa = 0.4036 g (0.4%) S No, 100 μL EtOH = 3.9054 g (4%)PEG 400 = 95.6 (95.6%) 100 Rapa = 2.0025 g (2%) S Yes, 1 μL, 3 μL, 20μL, EtOH = 3.98 g (4%) 40 μL PEG 400 = 94.00 g (94%) 101 Rapa = 9.5 mg(0.472%) S EtOH = 90.3 mg (4.485%) PEG 600 = 1913.5 mg (95.043%) 102Rapa = 44.6 mg (2.21%) S EtOH = 86.1.0 mg (4.26%) PEG 600 = 1891.1 mg(93.53%) 103 Rapa = 1.97 g (2%) S EtOH = 4.10 g (4%) PEG 400 = 94.15 g(94%) 104 Rapa = 1.95 g (2%) S EtOH = 4.00 g (4%) PEG 400 = 94.0 g (94%)105 Rapa = 8.00 g (2%) S PEG 400 = 376.0 g (94%) EtOH = 16.0 g (4%) 106Rapa = 6.00 g (2%) S PEG 400 = 282.0 g (94%) EtOH = 12.0 g (4%) 107 Rapa= 8.9 mg (0.4434%) S EtOH = 80.3 mg (4.0006%) PEG 300 = 1918.0 mg(95.556%) 108 Rapa = 40.8 mg (2.00886%) S EtOH = 110.0 mg (5.41605%) PEG300 = 1880.2 mg (92.57509%) 109 Rapa = 9.9 mg (0.488%) S EtOH = 86.7 mg(4.277%) PEG 400/300(50/50) = 1930.3 mg (95.235%) 110 Dexamethasone =142.5 mg (4.994%) SP 0.3305 μm Yes, 30 μL PEG 400 = 2710.7 mg (95.006%)111 Dexamethasone = 134.3 mg (4.891%) SP >10 μm PEG 400 = 2611.4 mg(95.109%) 112 Triamcinolone = 139.2 mg (5.087%) SP 3.98 μm Yes, 30 μLPEG 400 = 2597.4 mg (94.913%) 113 Triamcinolone = 135.3 mg (5.089%)SP >10 μm PEG 400 = 2523.5 mg (94.911%) 114 EtOH = 206.4 mg (4.121%) SNo, 30 μL PEG 400 = 4801.6 mg (95.879%) 115 Rapa = 43.0 mg (2.144%) SP61.4390 μm PEG 400 = 1962.3 mg (97.8567%) 116 Rapa = 40.0 mg (2.001%) SP3.7128 μm PEG 400 = 1959.1 mg (97.999%) 117 Rapa = 42.9 mg (2.142%) SP2.7313 μm PEG 400 = 1959.7 mg (97.858%) 118 Rapa = 100.8 mg (2.013%) SP4.1063 μm PEG 400 = 4906.0 mg (97.987%) 119 Rapa = 20.9 mg (0.42%) SEtOH = 209.1 mg (4.17%) PEG 400 = 4784.9 mg (95.41%) 120 Rapa = 20.6 mg(0.41%) S EtOH = 211.5 mg (4.22%) Benz. Chl = 19.1 mg (0.38%) PEG 400 =4762.0 mg (94.99%) 121 Rapa = 20.1 mg (0.40%) S EtOH = 211.5 mg (4.22%)Benz. Chl = 2.3 mg (0.05%) PEG 400 = 4782.3 mg (95.34%) 122 Rapa = 8.0 g(2%) S EtOH = 16.0 g (4%) PEG 400 = 376.0 g (94%) 123 Rapa = 351.3 mg(2.006%) S EtOH = 2353.1 mg (4.093%) PEG 400 = 16448.2 mg (93.901%) 124Rapa = 2.2035 g (2%) S EtOH = 4.45 g (4%) PEG 400 = 103.7 g (94%) 125Rapa = 515.5 mg (2.021%) SP 18.1453 μm PEG 400 = 24,993.8 mg (97.979%)126 Rapa = 0.3 g (2%) S EtOH = 0.6 g (4%) PEG 400 = 14.1 g (94%) BHT =0.0002 (0.002%) 127 Rapa = 0.3 g (2%) S EtOH = 0.6 g (4%) PEG 400 = 14.1g (94%) BHT = 0.00037 (0.004%) 128 Rapa = 0.3 g (2%) S EtOH = 0.6 g (4%)PEG 400 = 14.1 g (94%) BHT = 0.0081 (0.05%) 129 Rapa = 243.2 mg (1.869%)S EtOH = 4.88.4 mg (3.753%) PEG 400 = 12283.3 mg (94.378%) 130 Rapa =0.404 g (2%) S EtOH = 0.8 g (4%) PEG 400 = 18.8 g (94%) BHT = 0.00051(0.002%) 131 Rapa = 0.6024 g (2%) S EtOH = 1.2 g (4%) PEG 400 = 28.25 g(94%) 132 Rapa = 2.001 g (2%) S EtOH = 4.05 g (4%) PEG 400 = 94.45 g(94%) 133 Rapa = 0.5155 g (2.057%) S EtOH = 1.0198 g (4.070%) PEG 400 =23.5225 g (93.873%) 134 PEG 400 = 9.6 g (96%) S EtOH = 0.4 g (4%) 135Rapa = 0.610 g (2%) S EtOH = 1.2 g (4%) PEG 400 = 28.2 g (94%) 136 Rapa= 24.6 mg (1.193%) S EtOH = 91.1 mg (4.418%) Tyloxapol = 219.6 mg(10.649%) BSS = 1726.8 mg (83.740%) 137 Rapa = 100.0 mg (1.993%) SP PEG400 = 4916.9 mg (98.007%) 138 Rapa = 201.6 mg (4.005%) SP PEG 400 =4831.5 mg (95.995%) 139 Rapa = 102.4 mg (2.036%) S EtOH = 209.0 mg(4.154%) PEG 400 = 4719.3 mg (93.810%) 140 Rapa = 10.3 mg (0.205%) SYes, 10 μL EtOH = 27.4 mg (0.544%) PEG 400 = 4995.8 mg (99.251%) 141Rapa = 10.6 mg (0.211%) S No, 10 μL EtOH = 208.4 mg (4.150%) PEG 400 =4802.3 mg (95.639%) 142 Rapa = 31.5 mg (0.628%) S Yes, 10 μL EtOH = 67.1mg (1.337%) PEG 400 = 4918.9 mg (98.035%) 143 Rapa = 30.8 mg (0.613%) SNo, 10 μL, 100 μL EtOH = 204.5 mg (4.073%) PEG 400 = 4786.1 mg (95.314%)144 Rapa = 103.5 mg (2.057%) S Yes, 10 μL EtOH = 207.1 mg (4.116%) PEG400 = 4720.8 mg (93.827%) 145 Rapa = 283.0 mg (2.020%) S EtOH = 566.1 mg(4.041%) PEG 400 = 13,160.8 mg (93.939%) 146 Rapa = 280.1 mg (1.998%) SEtOH = 565.2 mg (4.033%) PEG 400 = 13,171.7 mg (93.969%) 147 Rapa =201.6 mg (3.000%) SP PEG 400 = 6518.8 mg (97.000%) 148 Rapa = 31.9 mg(1.019%) S Benzyl Alcohol = 1021.9 mg (20.070%) Sesame Oil = 4017.9 mg(78.911%) 149 Rapa = 51.5 mg (1.03%) S Benzyl Alcohol = 259.9 mg (5.19%)Sesame Oil = 4694.3 mg (93.78%) 150 Rapa = 5.96 g (2%) S EtOH = 12.0 g(4%) PEG 400 = 282.0 g (94%) 151 Rapa = 54.5 mg (1.07%) S Benzyl Alcohol= 1014.3 mg (19.95%) Olive Oil = 4014.8 mg (78.98%) 152 Rapa = 0 mg(0.00%) S Benzyl Alcohol = 269.4 mg (5.421%) Tyloxapol = 608.2 mg(12.238%) Sesame Oil = 4092.2 mg (82.341%) 153 Rapa = 76.3 mg (1.75%) SBenzyl Alcohol = 307.0 mg (7.06%) Tyloxapol = 607.8 mg (13.97%) SesameOil = 3000.5 mg (68.97%) Span 80 = 63.1 mg (1.45%) EtOH = 295.5 mg(6.79%) 154 Form. # 150 = 200 g (99.998) S BHT = 0.004 g (0.002%) 155Rapa = 51.0 mg (0.87%) S EtOH = 642.3 mg (10.93%) Benzyl Alcohol = 431.8mg (7.34%) Sesame Oil = 4753.7 mg (80.86%) 156 Rapa = 51.4 mg (1.03%) SBenzyl Alcohol = 518.4 mg (10.34%) Olive Oil = 4444.7 mg (88.64%) 157Rapa = 8.1 g (2%) S EtOH = 16.0 g (4%) PEG 400 = 376.0 g (94%) 158 Form.# 157 = 225.00 g (99.998%) S BHT = 0.0045 g (0.002%) 159 Rapa = 8.1 g(2%) S EtOH = 16.0 g (4%) PEG 400 = 376 g (94%) 160 Form. # 159 = 112.0g (99.998%) S BHT = 0.00224 g (0.002%) 161 Form. # 159 = 112.0 g(99.998%) S BHT = 0.0019 g (0.002%) 162 Rapa = 55.4 mg (1.10%) S EtOH =112.7 mg (2.25%) Benzyl Alcohol = 157.8 mg (3.15%) Cotton Seed Oil =4688.0 mg (93.50%) 163 Rapa = 5.005 g (1%) S EtOH = 10.0 g (2%) PEG 400= 485.5 g (97%) 164 PEG 400 = 9.82 g (98%) S EtOH = 0.235 g (2%) 165Form. # 163 = 100.25 g (99.998%) S BHT = 0.0026 g (0.002%) 166 Rapa =203.1 mg (2.025%) SP 2.8651 μm F68 = 30.3 mg (0.303%) Sterile Water =9792.6 mg (97.672%) 167 Rapa = 201.4 mg (2.0005%) SP 1.0984 μm Tween 20= 43.9 mg (0.436%) Sterile Water = 9822.8 mg (97.564%) 168 EtOH = 0.8301g (4.144%) S PEG 400 = 19.2014 g (95.856%) 169 Form. # 168 = 300 μl S170 Form. # 168 = 250 μl S Form. # 154 = 50 μl 171 Form. # 168 = 200 μlS Form. # 154 = 100 μl 172 Form. # 168 = 150 μl S Form. # 154 = 150 μl173 Form. # 154 = 300 μl S 174 Rapa = 102.2 mg (2.041%) SP 0.4165 μm F68= 16.0 mg (0.32%) Sterile Water = 4889.0 mg (97.639%) 175 Rapa = 101.1mg (2.010%) SP 0.5294 μm Tween 20 = 27.7 mg (0.551%) Sterile Water =4901.0 mg (97.439%) 176 BSS+ = 0 μl S Sterile Water = 0 μl Form. # 154 =1000 μl 177 BSS+ = 200 μl SP Sterile Water = 0 μl Form. # 154 = 800 μl178 BSS+ = 400 μl SP Form. # 154 = 600 μl 179 BSS+ = 500 μl SP Form. #154 = 500 μl 180 BSS+ = 600 μl SP Form. # 154 = 400 μl 181 BSS+ = 800 μlSP Form. # 154 = 200 μl 182 Sterile Water = 200 μl SP Form. # 154 = 800μl 183 Sterile Water = 400 μl SP Form. # 154 = 600 μl 184 Sterile Water= 500 μl SP Form. # 154 = 500 μl 185 Sterile Water = 600 μl SP Form. #154 = 400 μl 186 Sterile Water = 800 μl SP Form. # 154 = 200 μl 187 BSS+= 2536.9 mg (49.98%) SP 60.2075 μm Form. # 154 = 2538.7 mg (50.02%) 188Sterile Water = 2515.6 mg (49.84%) SP 617.5157 μm Form. # 154 = 2532.2mg (50.16%) 189 F68 = 12.6 mg (0.25%) SP 70.6089 μm Sterile Water =2524.7 mg (49.79%) Form. # 154 = 2533.1 mg (49.96%) 190 Rapa = 2.0225 g(2%) S EtOH = 3.65 g (4%) PEG 400 = 94.0 g (94%) BHT = 0.002 g (0.002%)191 F68 = 12.1 mg SP Sterile Water = 2558.9 mg Form. # 154 = 2556.4 mg192 F68 = 19.8 mg SP Sterile Water = 2564.1 mg Form. # 154 = 25557.5 mg193 F68 = 25.3 mg SP Sterile Water = 2575.1 mg Form. # 154 = 2572.9 mg194 F68 = 32.4 mg SP Sterile Water = 2572.1 mg Form. # 154 = 2562.1 mg195 F68 = 38.3 mg SP Sterile Water = 2563.2 mg Form. # 154 = 2573.5 mg196 F68 = 43.6 mg SP Sterile Water = 2541.1 mg Form. # 154 = 2556.0 mg197 F68 = 51.2 mg SP Sterile Water = 2594.5 mg Form. # 154 = 2594.1 mg198 PEG 400 = 1920 g (96%) S EtOH = 80 g (4%) 199 Form. # 168 = 1000 μlS 200 Form. # 168 = 200 μl S Form. # 154 = 800 μl 201 Form. # 168 = 400μl S Form. # 154 = 600 μl 202 Form. # 168 = 500 μl S Form. # 154 = 500μl 203 Form. # 168 = 600 μl S Form. # 154 = 400 μl 204 Form. # 168 = 800μl S Form. # 154 = 200 μl 205 PEG 400 = 200 μl S Form. # 154 = 800 μl206 PEG 400 = 400 μl S Form. # 154 = 600 μl 207 PEG 400 = 500 μl S Form.# 154 = 500 μl 208 PEG 400 = 600 μl S Form. # 154 = 400 μl 209 PEG 400 =800 μl S Form. # 154 = 200 μl 210 Phosal 50PG = 6735.0 mg (99.002%) STween 80 = 67.9 mg (0.998%) 211 Rapa = 2.0047 g (2%) S EtOH = 4.00 g(4%) PEG 400 = 94.05 g (94%) 212 Phosal 50PG = 20.0662 g (98.999%) STween 80 = 0.2029 g (1.001%) 213 Form. # 154 = 100 μl S Form. # 168 =900 μl 214 Form. # 154 = 100 μl S Form. # 168 = 900 μl 215 Form. # 154 =100 μl S Form. # 168 = 900 μl 216 Form. # 154 = 100 μl S PEG 400 = 900μl 217 Form. # 154 = 100 μl S PEG 400 = 900 μl 218 Form. # 154 = 100 μlS PEG 400 = 900 μl 219 Form. # 154 = 100 μl SP BSS+ = 900 μl 220 Form. #154 = 100 μl SP BSS+ = 900 μl 221 Form. # 154 = 100 μl SP BSS+ = 900 μl222 Form. # 154 = 1000 μl S 223 Form. # 154 = 1000 μl S 224 Form. # 154= 100 μl S Form. # 168 = 900 μl 225 Form. # 154 = 100 μl S Form. # 168 =900 μl 226 Form. # 154 = 100 μl S Form. # 168 = 900 μl 227 Form. # 154 =100 μl S PEG 400 = 900 μl 228 Form. # 154 = 100 μl S PEG 400 = 900 μl229 Form. # 154 = 100 μl S PEG 400 = 900 μl 230 Form. # 154 = 100 μl SPBSS+ = 900 μl 231 Form. # 154 = 100 μl SP BSS+ = 900 μl 232 Form. # 154= 100 μl SP BSS+ = 900 μl 233 Form. # 154 = 200 μl S Form. # 168 = 800μl 234 Form. # 154 = 200 μl S Form. # 168 = 800 μl 235 Form. # 154 = 200μl S Form. # 168 = 800 μl 236 Form. # 154 = 200 μl S Form. # 168 = 800μl 237 Form. # 154 = 200 μl S PEG 400 = 800 μl 238 Form. # 154 = 200 μlS PEG 400 = 800 μl 239 Form. # 154 = 200 μl SP BSS+ = 800 μl 240 Form. #154 = 200 μl SP BSS+ = 800 μl 241 Form. # 154 = 200 μl SP BSS+ = 800 μl242 Form. # 154 = 100 μl S No, 10 μL Form. # 168 = 900 μl 243 Form. #154 = 100 μl S Yes, 10 μL PEG 400 = 900 μl 244 Form. # 154 = 100 μl SPYes, 10 μL BSS+ = 900 μl 245 Form. # 154 = 100 μl SP BSS+/CMC(0.5%) =900 μl 246 Form. # 154 = 400 μl S No, 10 μL Form. # 168 = 900 μl 247Form. # 154 = 400 μl S Yes, 10 μL PEG 400 = 900 μl 248 Form. # 154 = 400μl SP Yes, 10 μL BSS+ = 900 μl 249 Form. # 154 = 400 μl SPBSS+/CMC(0.5%) = 900 μl 250 Form. # 154 = 100 μl SP BSS+/CMC(0.5%) = 900μl 251 Form. # 154 = 100 μl SP BSS+/CMC(0.5%) = 900 μl 252 Form. # 154 =100 μl SP BSS+/CMC(0.5%) = 900 μl 253 Form. # 154 = 200 μl SPBSS+/CMC(0.5%) = 800 μl 254 Form. # 154 = 200 μl SP BSS+/CMC(0.5%) = 800μl 255 Form. # 154 = 200 μl SP BSS+/CMC(0.5%) = 800 μl 256 Form. # 154 =400 μl SP BSS+/CMC(0.5%) = 900 μl 257 Form. # 154 = 400 μl SPBSS+/CMC(0.5%) = 900 μl 258 Form. # 154 = 400 μl SP BSS+/CMC(0.5%) = 900μl 259 EtOH = 17.1 mg (0.57%) S PEG 400 = 2997.3 mg (99.43%) 260 EtOH =40.8 mg (1.35%) S PEG 400 = 2980.2 mg (98.65%) 261 EtOH = 47.1 mg(1.57%) S PEG 400 = 2950.1 mg (98.43%) 262 Rapa = 2.0032 g (2%) S EtOH =3.92 g (4%) PEG 400 = 94.00 g (94%) 263 Triamcinolone acetomide = 80.8mg (4.04%) SP PEG 400 = 1920.8 mg (95.96%) 264 NFF-0007 filled in glovebox S 265 PEG 400 = 9.598 g (96%) S EtOH = 0.4052 (4%) 266 Triamcinoloneacetomide = 42.2 mg SP (4.123%) PEG 400 = 981.3 mg (95.877%) 267 Phosal50PG = 20.0783 g (99.00835%) S Tween 80 = 0.2011 g (0.99165%) 268 PEG400 = 96.1 g (96%) S EtOH = 4.00 g (4%) 269 Rapa = 0.4001 g (2%) S EtOH= 0.80 g (4%) PEG 400 = 18.8 g (94%) 270 Sterile Water = 9955.8 mg(99.27%) S CMC High visc. = 47.8 mg (0.48%) Tween 80 = 25.4 mg (0.25%)271 Sterile Water = 9947.5 mg (99.00%) S CMC Medium visc. = 75 mg(0.75%) Tween 80 = 25.1 mg (0.25%) 272 Rapa = 41 mg (2.01%) SP Form. #270 = 2000 mg (97.99%) 273 Rapa = 40.2 mg (1.97%) SP MSF-03-172-07E =2000 mg (98.03%) 274 NMP (Pharmasolve ®⁾ = 1280.5 mg (65.89%) S PLGA75/25 = 662.9 mg (34.11%) 275 NMP (Pharmasolve ®⁾ = 1573.3 mg (80.50%) SPLGA 75/25 = 381.0 mg (19.50%) 276 NMP (Pharmasolve ®⁾ = 1009.7 mg49.8%) S Yes, 10 μL PLGA 75/25 = 1001.6 mg (50.20%) 277 Sterile Water =14934.0 mg (99.25%) S CMC Medium visc. = 112.4 mg (0.75%) 278 PropyleneGlycol = 1893.7 mg (93.85%) S Yes, 10 μL EtOH = 83.8 mg (4.16%) Rapa =40.2 mg (1.99%) 279 Propylene Glycol = 1946.2 mg (95.68%) S Yes, 10 μLBenzyl Alcohol = 47.1 mg (2.31%) Rapa = 40.8 mg (2.01%) 280 PEG 300 =1894.1 mg (93.74%) S Yes, 10 μL EtOH = 40.1 mg (1.98%) Rapa = 86.4 mg(4.28%) 281 PEG 300 = 1925.5 mg (95.88%) S Yes, 10 μL, 30 μL EtOH = 39.8mg (1.98%) Rapa = 43.0 mg (2.14%) 282 Rapa = 100.6 mg (2.01%) SP Yes, 10μL, 30 μL MSF-03-176-02 = 4910.8 mg (97.99%) 283 Rapa = 11.5 mg (0.57%)S PEG 300 = 2012.5 mg (99.43%) 284 Rapa = 10.3 mg (0.51%) S PEG 400 =2017.2 mg (99.49%) 285 Rapa = 9.8 mg (0.486%) S PEG 600 = 2005.9 mg(99.51%) 286 Tacrolimus = 42.7 mg (2.11%) S EtOH = 46.0 mg (2.27%) PG =1938.7 mg (95.62%) 287 Tacrolimus = 40.7 mg (2.01%) S EtOH = 43.0 mg(2.12%) PEG 300 = 1942.1 mg (95.87%) 288 Tacrolimus = 40.3 mg (1.99%) SEtOH = 43.8 mg (2.16%) PEG 400 = 1942.3 mg (95.85%) 289 Tacrolimus =40.8 mg (2.03%) S EtOH = 44.5 mg (2.21%) PEG 600 = 1924.0 mg (95.76%)290 Rapa = 61.0 mg (3.17%) S NMP = 1226.54 mg (63.80%) PLGA 75/25 =634.96 mg (33.03%) 291 Rapa = 100.2 mg (5.13%) S NMP = 1492.95 mg(76.37%) PLGA 75/25 = 361.65 mg (18.50%) 292 Rapa = 62.9 mg (3.04%) SNMP = 1103.8 g mg (53.40%) PLGA 75/25 = 900.2 mg (43.56%) 293 Rapa =62.4 mg (3.00%) S NMP = 1205.1 mg (58.11%) PLGA 75/25 = 806.4 mg(38.89%) 294 Water + 1% CMC Med. = 4909.1 mg SP (97.99%) Rapa = 100.5 mg(2.01%) 295 Water + 1% CMC high. = 4903.8 mg (97.96%) SP Rapa = 101.9 mg(2.04%) 296 Rapa = 40.5 mg (2.03%) S NMP = 1958.7 mg (97.97%) 297 Rapa =20.5 mg (2.0%) SP DMA = 41.4 mg (4.0%) PVP = 35.0 mg (3.4%) H2O = 934.7mg (90.6%) 298 Rapa = 10.6 mg (2.0%) S DMA = 10.6 mg (2.0%) PEG 400 =506.1 mg (96%) 299 Rapa = 5.2 mg (2.0%) SP 1% DMA in PEG 400 = 257.4 mg(98%) 300 Rapa = 20.0 mg (2.0%) S DMA = 7.8 mg (0.8%) PEG 400 = 974 mg(97.2%) 301 Rapa = 20.1 mg (1.3%) S DMA = 19.5 mg (1.3%) PEG 400 =1449.6 mg (97.3%) 302 Rapa = 20.0 mg (2.0%) SP PVP = 10.8 mg (1.1%) PEG400 = 994.5 mg (97.0%) 303 Rapa = 20.4 mg (2.0%) SP PVP = 24.5 mg (2.4%)PEG 400 = 990.7 mg (95.7%) 304 Rapa = 25.5 mg (2.4%) SP PVP = 51.9 mg(4.8%) PEG 400 = 1000.6 mg (92.8%) 305 Rapa = 22.5 mg (2.3%) S BA = 27.5mg (2.7%) PEG 400 = 950.7 mg (95.0%) 306 Rapa = 30.2 mg (2.3%) SP PVP =240.9 mg (18.6%) PEG 400 = 1021.2 mg (79.0%) 307 Rapa = 8.7 mg (3.1%) SP1% PVP in H2O = 273 mg (96.9%) 308 Rapa = 12.6 mg (2.53%) SP 5% PVP inH2O = 501.6 mg (97.5%) 309 Rapa = 20.3 mg (3.8%) SP 10% PVP in H2O =513.9 mg (96.2%) 310 Rapa = 100.5 mg (2.0%) S Yes, 10 μL DMA = 67.8 mg(1.4%) PEG 400 = 4838.3 mg (96.6%) 311 Rapa = 96.8 mg (1.9%) S Yes, 10μL BA = 157.5 mg (3.2%) PEG 400 = 4748.7 mg (94.9%) 312 Rapa = 105.8 mg(2.1%) S DMA = 5.63 mg (0.1%) PEG 400 = 4888.9 mg (97.8%) 313 Rapa =20.2 mg (2.0%) SP PVP = 99.2 mg (9.9%) H2O = 882.3 mg (88.1%) 314 Rapa =100.3 mg (2.0%) SP PVP = 251.4 mg (5.0%) H2O = 4662.8 mg (93.0%) 315Rapa = 20.3 mg (2.0%) S DMA = 983.9 mg (98%) 316 Triamcinolone = 22.8 mg(2.0%) S Yes, 10 μL DMA = 12.0 mg (1.1%) PEG 400 = 1104.5 mg (96.9%) 317Triamcinolone = 1.0 mg (0.1%) S EtOH = 49.30 mg (4.0%) PEG 400 = 1191.9mg (96.0%) 318 Triamcinolone = 18.7 mg (0.9%) S PEG 400 = 959.8 mg(99.1%) 319 Triamcinolone = 25.5 mg (1.3%) S EtOH = 83.0 mg (4.1%) PEG400 = 1905.6 mg (94.6%) 320 Dexamethasone = 20.4 mg (1.2%) S EtOH = 71.7mg (4.1%) PEG 400 = 1737.6 mg (98.8%) 321 Dexamethasone = 27.5 mg (2.0%)S Yes, 10 μL DMA = 5.6 mg (0.4%) PEG 400 = 1347.3 mg (97.6%) 322 Rapa =9.1 mg (0.152%) E EtOH = 90.9 mg (1.514%) F127 = 262.8 mg (4.378%) Water= 1489.1 mg (24.804%) Sesame oil = 4151.5 mg (69.152%) 323 Rapa = 24.4mg (0.625%) E Phosal 50PG = 203.1 mg (5.201%) EtOH = 166.8 mg (4.272%)Labrafac CC = 1502.8 mg (38.486%) Sesame oil = 2007.7 mg (51.416%) 324Form. # 174 with 2 mm beads SP 0.4929 μm 325 Form. # 175 with 2 mm beadsSP 0.4804 μm 326 Rapa = 4 mg (4%) S Ethanol = 4 mg (4%) PEG 400 = 92 mg(92%) 327 Rapa = 1.47% S Ethanol = 2.93% PEG 400 = 69% Saline = 26.6%328 Rapa = 1.38% SP Ethanol = 2.75% PEG 400 = 64.62% Saline = 31.25% 329Rapa = 0.07% E Cremophor (PEG 35 castor oil) = 8.35% Capmul MCM (C8)(mono/diglycerides of caprylic acid) = 8.37% Sterile Water = 83.21% 330Rapa = 0.09% E Tyloxapol (nonionic polymer of the alkyl aryl polyetheralcohol) = 8.12% Capmul MCM (C8) (mono/diglycerides of caprylic acid) =8.15% Ethanol = 0.35% Sterile Water = 83.29% 331 Rapa = 0.08% ETyloxapol (nonionic polymer of the alkyl aryl polyether alcohol) = 6.78%Capmul MCM (C8) (mono/diglycerides of caprylic acid) = 6.81% Phosal ®50PG = 3.12% Sterile Water = 83.21% 332 Rapa = 0.17% E Cremophor (PEG 35castor oil) = 7.18% Capmul MCM (C8) (mono/diglycerides of caprylic acid)= 9.47% Sterile Water = 83.18% 333 N-methyl Pyrrolidone (NMP) = 10.5% SWater = up to 15.8% Propylene Glycol (PG) = 73.7% Rapamycin = up to 5%(w/w) 334 N-methyl Pyrrolidone (NMP) = 38.9% S Water = up to 22.2%Propylene Glycol (PG) = 38.9% Rapamycin = up to 5% (w/w) 335 N-methylPyrrolidone (NMP) = 10.5% S Water = up to 15.8% PEG 600 = 73.7%Rapamycin = up to 5% (w/w) 336 Dimethyl Acetamine (DMA) = 10.5% S Water= up to 15.8% Propylene Glycol (PG) = 73.7% Rapamycin = up to 5% (w/w)337 Dimethyl Sulfoxide (DMSO) = 10.5% S Water = up to 15.8% PropyleneGlycol (PG) = 73.7% Rapamycin = up to 5% (w/w) 338 N-methyl Pyrrolidone(NMP) = 27.9% S Water = up to 28.4% PEG 400 = 39% Ethanol = 4.7%Rapamycin = up to 5% (w/w) 339 Dasatanib = 4.84% S Dimethyl Acetamine(DMA) = 15% PEG 400 = 80.16% 340 Rapa = 40 mg (2%) S Ethanol = 80 mg(4%) PEG 400 = 1880 mg (94%) 341 Rapa = 0.3% Tyloxapol = 4.8%Lauroglycol = 2.8% Labrafac = 1.8% remainder Sterile Water 342 Rapa =0.2% Tyloxapol = 2.8% EtOH = 0.3% Labrafac = 2.8% remainder SterileWater 343 Rapa = 0.2% Tyloxapol = 2.8% EtOH = 0.3% Labrafac = 2.8%remainer Sterile Water 344 Rapa = 0.2% EtOH = 0.4% PEG 400 = 69.4%remainder Sterile Water 345 Rapa = 0.2% EtOH = 0.4% PEG 400 = 69.2%remainder Sterile Water 346 Rapa = 0.6% (0.4% degradation products) SN-methyl pyrrolidone (NMP) = 30% Propylene Glycol (PG) = 39% remainderSterile Water 347 Rapa = 2.1% SP 3.5 μm CMC = 1% Sterile Water = 96.9%348 Rapa = 2.1% ISG PLGA 75/25 = 21.1% N-methyl pyrrolidone (NMP) =76.8% 349 Rapa = 2.0% N-methyl pyrrolidone (NMP) = 44.5% PEG 400 = 29.0%Water = 24.5% 350 Rapa = 1% Ethanol = 4% PEG 400 = 95% 351 Rapa = 10.0%SDF PVP 90 = 57.9% Phosal 50 SD = 20.0% PEG 400 = 10.1% Gamma Tocopherol= 1.0% Ascorbyl palmitate = 1.0% 352 Dexamethasone = 25.3% SDF HPC =33.2% Eudragit RS 30D = 32.0% Xanthan Gum = 9.5% Depending on theirtype, the listed formulations are denoted as one or more of solutions(S), suspensions (SP), emulsions (E), in situ gels (ISG) or solid dosageforms (SDF).

TABLE 2 Aqueous Humor Rapa Concentration 2% Rapa-PEG-EtOH Solution Mean[Rapa] (ng/mL) SD (ng/mL)  1.0 μL intravitreal 0.438 (1 day afterinjection) 0.141  3.0 μL intravitreal 0.355 (1 day after injection)0.234  3.0 μL sub-conj 0.338 (1 day after injection) 0.122  5.0 μL intoanterior chamber 0.167 (14 days after 0.183 injection) 10.0 μL intoanterior chamber 0.004 (14 days after 0.006 injection)

TABLE 3 Topical Administration Rapamycin Levels Cornea, ng/g Blood, Gr.II ng/ml Gr. I OD OS Gr. III time, day Gr. I Gr. II Gr. III (n = 2) std(n = 1) (n = 1) (n = 2) std 1 4.63 2.84 121.72 2.22 147.26 4.10 2 65.724.29 229.49 10.03 209.98 10.46 3 37.14 27.29 173.76 20.17 184.05 11.94 815.54 168.75 27.93 10 7.67 110.10 42.14 12 2.88 97.90 0.71 14 48.3284.20 31.25

TABLE 4 Topical Administration Rapamycin Levels R/Ch, ng/g time, Group IGroup II Group III day (n = 2) std OD (n = 1) OS (n = 1) (n = 2) std 120.23 0.0354 34.51 11.58 2 41.53 4.5679 64.86 25.68 3 48.92 36.069522.88 63.67 8 107.24 80.5748 10 139.98 150.1895 12 30.05 12.7845 14117.50 116.9555

TABLE 5 Topical Administration Rapamycin Levels Sclera, ng/g time, GroupI Group II Group III day (n = 2) std OD (n = 1) OS (n = 1) (n = 2) std 190.80 40.59 420.60 34.20 2 751.35 471.29 1353.60 144.50 3 350.80 77.22540.60 104.20 8 895.20 368.83 10 612.65 530.97 12 130.85 60.17 14 419.65515.41

TABLE 6 Topical Administration Rapamycin Levels Vitreous, ng/ml time,Gr. I Gr. II Gr. III day (n = 2) std OD (n = 1) OS (n = 1) (n = 2) std 11.24 0.1697 2.06 1.00 2 5.41 1.2799 5.01 2.95 3 3.03 0.6859 3.96 4.00 819.69 22.5072 10 7.90 7.2903 12 1.46 0.5374 14 5.66 2.6870

What is claimed is:
 1. A method for treating wet age-related maculardegeneration in a human subject by repeated administration of a firstformulation and a second formulation, the method comprising: (a)administering to the human subject two or more doses of a volume of thefirst formulation by intravitreal placement, wherein the volume of thefirst formulation comprises an amount of and about 500 μg ofranibizumab; and (b) administering to the human subject two or moredoses of a volume of the second formulation by intravitreal orsubconjuctival placement, wherein the volume of the second formulationcomprises an amount of between about 200 μg and about 2000 μg ofrapamycin, or a pharmaceutically acceptable salt or ester thereof;wherein the administration of rapamycin or a pharmaceutically acceptablesalt or ester thereof decreases the frequency at which ranibizumab isadministered to treat wet age-related macular degeneration in the humansubject.
 2. The method of claim 1, wherein step (a) is coincident withstep (b).
 3. The method of claim 1, wherein step (b) is subsequent tostep (a).
 4. The method of claim 1, wherein step (b) is prior to step(a).
 5. The method of claim 1, wherein the volume of the secondformulation contains a rapamycin dose selected from the group consistingof about 220 μg, about 440 μg, about 880 μg, and about 1320 μg ofrapamycin.
 6. The method of claim 1, wherein the second formulation is aliquid solution that consists essentially of about 2% (w/w) rapamycin,about 4% (w/w) ethanol, and about 94% (w/w) polyethylene glycol 400 (PEG400).
 7. The method of claim 1, wherein the second formulation is aliquid solution that consists essentially of about 4% (w/w) rapamycin,about 4% (w/w) ethanol, and about 92% (w/w) polyethylene glycol 400 (PEG400).
 8. The method of claim 1, wherein the second formulation is aliquid solution that consists essentially of about 1% (w/w) rapamycin,about 4% (w/w) ethanol, and about 95% (w/w) polyethylene glycol 400 (PEG400).
 9. The method of claim 1, wherein the second formulation is asolution of rapamycin dissolved in a solvent system.
 10. The method ofclaim 9, wherein the solvent system comprises polyethylene glycol. 11.The method of claim 10, wherein the solvent system further comprisesethanol.